LOCUS       Exported                9466 bp DNA     circular SYN 13-SEP-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9466)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 13, 2021 from SnapGene 5.3.2
            https://www.snapgene.com
COMMENT     Homologous recombination reaction between [pTIT.txt] as a target 
            sequence  and [PCR(N.xdna] as an inserted sequence.
FEATURES             Location/Qualifiers
     source          1..9466
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     148..164
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        172..190
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    695..7081
                     /label=L gene
     CDS             2342..2353
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      7152..7199
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(7222..7238)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(7275..7293)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7314..7330)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7338..7354
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(7362..7392)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7407..7428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7716..8304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8475..9335)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(9336..9440)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tttcgcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
       61 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      121 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgcgcgc gtaatacgac
      181 tcactatagg gcgaattaat tccggttatt ttccaccata ttgccgtctt ttggcaatgt
      241 gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct
      301 cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc
      361 ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga
      421 caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc
      481 ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt
      541 attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg
      601 gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaacgtct aggccccccg
      661 aaccacgggg acgtggtttt cctttgaaaa acacatgttg atcgattcag gagaagttta
      721 tgacgacccg gtggatccaa ttgagtcaga ggccgaacct agaggatctc ccactgtccc
      781 caacatcctg aggaactctg actataatct caactcccct ctaatagaag atcctgccag
      841 actgatgtta gaatggttaa agacagggaa tagaccttac cggatgacct taacagacaa
      901 ctgctctagg tcttataaag tctttaaaga ttacttcaag aagttagatt tgggatctgt
      961 taaagtaggt ggaactgcag cccagtcgat gatctctctc tggttgtatg gtgcccactc
     1021 tgaatcaaac aggagccgga gatgcatagc ggacttggct catttctatt ccaagtcttc
     1081 acccatagag aagttgttaa attgtacact tgggaataga ggactaagaa ttcccccaga
     1141 aggggtgttg agttgccttg aaagaattga ctatgataaa gcattcggga ggtatcttgc
     1201 caatacatat tcctcttatt tgtttttcca cgtgattacc ctgtacatga atgccctgga
     1261 ctgggaggaa gagaagacta ttttagcttt atggaaagat ttgacttcag tcgatattgg
     1321 aaaagattta gtcaagttca aagatcaaat atgggggttg ttaattgtga cgaaggactt
     1381 tgtgtactct cagagttcaa gctgtctttt tgacaggaac tatacactca tgctgaaaga
     1441 cctttttttg tctcgcttca actctttgat gattctgctt tctccccctg aaccccgata
     1501 ttcagacgac ctgatatctc agctatgcca gctgtatatt gccggggatc aagtattgtc
     1561 tatgtgtgga aactccggtt atgaagtcat taaaatactg gaaccatatg tcgtgaacag
     1621 cttagtccag agggcagaga agttcaggcc tctcatccat tctttgggag atttccctgt
     1681 gtttatcaaa gacaaggtga gtcaacttga gggaacattc ggtcctagtg caaaaaggtt
     1741 ttttagagtt ctggatcagt tcgacaacat acatgacctg gtctttgtat atggctgtta
     1801 caggcattgg ggacatcctt atatagatta tagaaagggt ctgtcgaaac tatatgatca
     1861 agttcatatc aagaaggtga tagataagtc ctaccaggag tgcttggcga gcgaccttgc
     1921 ccgaaggatc ctcaggtggg gatttgacaa gtactccaaa tggtacattg actctcgatt
     1981 tcttcctaga gatcatccct tgactccgta catcaagact cagacatggc cacccaaaca
     2041 tatagtggat ttagtagggg acacatggca caagctccca atcacacaga tatttgagat
     2101 ccctgagtca atggacccat cagaaatact agatgacaag tcacactcct tcaccagaac
     2161 gaggctagcg tcttggttgt cagagaacag aggggggccc gttcccagtg aaaaggtcat
     2221 catcacagct ctgtctaaac cacctgtcaa tccgagggag tttttaaaat ctatcgacct
     2281 tggaggattg cctgatgaag acttgataat tggcctcaag ccgaaggaga gagagctgaa
     2341 gatcgagggt cgattttttg ccttgatgtc ttggaaccta agattgtact ttgttatcac
     2401 cgaaaagctc ttagctaatt atatcttgcc actctttgat gcactgacta tgacagacaa
     2461 tctgaacaag gtgttcaaga agttgattga cagagtgact gggcaagggc ttttggacta
     2521 ctcaagggtt acatatgcct ttcacctgga ctatgagaag tggaacaatc atcaaaggtt
     2581 ggagtcaaca gaagatgtat tttctgtcct ggatcaggtg tttggattaa aaagggtgtt
     2641 ttctaggact catgagtttt ttcagaagtc ctggatctat tattcagata gatcagacct
     2701 catcggtcta tgggaggacc aaatatactg cttagatatg tccaatggcc cgacctgttg
     2761 gaatggtcaa gatggagggc tagagggctt gcggcagaag ggctggagtc tggttagcct
     2821 attaatgata gacagagagt ctcaaaccag aaacacaaga accaaaatat tagctcaagg
     2881 agataaccag gttctatgtc caacatatat gttgtcacct gggctatcca gagagggact
     2941 tctctatgag ttggagagca tatcaagaaa tgcgctttca atataccgag ctatcgaaga
     3001 gggggcatcc aaactagggt tgattatcaa gaaggaagag accatgtgta gttacgactt
     3061 tctcatctat gggaagaccc ctttgtttag gggcaatatc ctagtacctg agtcaaaaag
     3121 atgggccaga gtctcttgca tatcaaacga tcaaatagtt aatcttgcca acataatgtc
     3181 aacggtgtcc acaaatgcat taactgtggc acaacactcc cagtccttga tcaagccaat
     3241 gagggacttt ctgcttatgt cagtccaggc agtttttcac tatttgttgt tcagcccgat
     3301 cctaaaaggc agagtttaca agattttaag tgctgaaggt gacaatttcc ttttagctat
     3361 gtcaagaata atctatttag acccatcctt ggggggagtg tctgggatgt ccctggggag
     3421 gttccatata cgtcagttct cggatcctgt ctctgagggg ttgtcttttt ggagagagat
     3481 ctggctgagc tcccatgagt cttggattca cgccttgtgt caggaggcgg ggaatccaga
     3541 tcttggagag agaacactcg agagcttcac tcggcttcta gaggacccca ccaccttaaa
     3601 tatcaaggga ggggcgagtc ccaccatctt actcaaggac gctatcagaa aagccctgta
     3661 tgatgaagtg gacaaggtgg agaactcaga gtttagagag gcaatcctct tgtctaagac
     3721 ccatagggat aactttatac ttttcttgaa atctgttgaa cccctatttc cccgatttct
     3781 cagtgagctc ttcagctcat ccttcttggg gatccctgag tcaatcattg gactgataca
     3841 aaactctcgg acgataagaa ggcagtttag aagaagtctc tcacgaactt tggaggagtc
     3901 tttttacaac tcggagatcc atggaatcaa caggatgacc cagacacctc aacgagtcgg
     3961 gagggtatgg ccttgctcct cggaaagggc agatctcttg agggaaatct cttgggggag
     4021 gaaagtggta ggtaccacgg tgcctcaccc ttctgagatg ttgggcctac ttcctaggtc
     4081 ttccatctct tgcacttgtg gaaccacagg aggagaaaat cccagagttt cagtatctgt
     4141 gctcccatcc tttgatcaat cgtttttctc acgaggcccc ctaaaggggt atttgggctc
     4201 atcaacctcc atgtctactc agctatttca tgcgtgggaa aaggtcacca atgttcatgt
     4261 ggtgaagagg gccctgtcac taaaagaatc tataaactgg ttcatcacaa gaaactctaa
     4321 tttggcgcag actctaatta ggaacataat gtctctgaca ggccctgatt tccctctgga
     4381 ggaggctcct gttttcaaaa ggacagggtc tgctttgcat agattcaagt ctgctagata
     4441 cagtgaaggg ggatattctt ctgtatgccc caatcttctc tctcatatct ccgtaagtac
     4501 agatactatg tctgacttga cccaggacgg aaagaactat gatttcatgt tccagccctt
     4561 gatgctttat gcgcaaacat ggacatctga actagtacaa agagacacga gactgaagga
     4621 ttcaacgttt cactggcacc tccgatgtaa caggtgtgta aggcccatcg acgatatcat
     4681 attggagacc tctcagatat tcgagtttcc ggacgtgtca aaaaggatat ctaggatggt
     4741 ttctggagct gtacctcatt ttcagaaact tcctgatatc cgtctgaaac caggagattt
     4801 tgaatctcta agtggcaggg agaaatctcg tcacataggt tcggctcaag ggctcttata
     4861 ttcgatcttg gttgcaatcc atgattcagg atataatgac agcaccatct tccctgtcaa
     4921 catatatagc aaggtttccc ccagagacta tttaagaggg cttgcaagag gggtcctgat
     4981 agggtcgtcg atctgtttct tgaccagaat gaccaacatc aacataaata gacctcttga
     5041 gttgatctca ggcgtgatct catatatcct cttgagattg gataatcacc catctttata
     5101 cataatgctc agagagccgt ctcttagagg agagatattt tcaatacccc aaaaaattcc
     5161 agcagcttac ccgaccacga tgaaagaggg caatcgatcc attttgtgtt acctccaaca
     5221 tgttctgcga tatgagcgag aggttatcac agcatcccca gagaatgatt ggttgtggat
     5281 cttctcggac ttcagaagtt ctaaaatgac gtacctaacc ctcattactt atcagtctca
     5341 tcttctactc cagagggttg aaagaaatct gtcgaagagc atgagaacta acctgcgaca
     5401 gatgagttcc ctgatgaggc aagtactagg agggcatgga gaggacacct tagaatctga
     5461 cgatgacatt caaaggctat tgaaggattc tctgcgcaga acgaggtggg tggaccaaga
     5521 agtgcgccat gcagccagaa ccatgacagg gggttacagt cctaacaaga aggtgtcgcg
     5581 caaggcgggg tgttcagagt gggtttgctc tgctcagcag gttgcagtct ccacctcagc
     5641 caacccagcc cctgtctcag agcttgatat aagggccctc tccaagaagt tccaaaaccc
     5701 tcttatctcg ggcttgagag ttgtccagtg ggcaactggc gcccattata agctcaagcc
     5761 cattctagac gatctcaatg ttttcccatc cctatgtcta gtggtaggag atggatcggg
     5821 gggaatatcg agagcagtcc tcaacatgtt tccggacgcc aaacttgtat tcaacagctt
     5881 gttggaggta aatgacctga tggcgtctgg gacacatcca ttgcccccct cagcaatcat
     5941 gagtggcggc gatgacatta tctccagagt agtggacttt gaatcaatct gggaaaagcc
     6001 gtcagatctg aggaacttga ccacctggaa gtacttccag tcagtccaaa aacaggtgaa
     6061 catgtcttat gacctaatca tttgtgacgc agaagtcact gacattgcat caattaatcg
     6121 gataacactg ttgatgtcag actttgcatt atccatagat ggtcctctct atctggtctt
     6181 caaaacttac ggaaccatgc tagtgaatcc ggaatataga gcgattcaac acttatcgag
     6241 agcgttccct tcagtcacag gatttataac ccaaatgacc tcgtcttttt catccgagct
     6301 gtacctcaga ttctccaagc gagggaagtt tttccgagat gctgagtact taacttcttc
     6361 tacccttcga gagatgagcc ttgttttatt caactgtagc agccccaaga gtgagatgca
     6421 gagggctcgc tctttgaatt accaagatct tgtaagagga ttccctgagg agattatatc
     6481 taacccttac aatgagatga tcataaccct gattgacagt gatgtggagt ccttcctggt
     6541 tcacaagatg gtggatgatc tggaattaca gagaggaacc ttgtctaaaa tgtctatcat
     6601 tatagccatt atgatagttt tctctaatag agtttttaat gtctccaaac cattgactga
     6661 tcccttgttc tatccaccgt ctgaccccaa gatcttgagg cacttcaata tctgctgcag
     6721 tactatgatg tatctatcta tggctttagg tgatgtcccc agctttgcaa ggcttcatga
     6781 cttgtacaat agacctataa cttattattt caaaaagcaa gtcattcgag ggaatattta
     6841 cctatcctgg agttggtctg atgacacctc agtattcaag agggtggcct gcaactctag
     6901 cttgagtctg tcatctcact ggatcaggct gatttacaag atagtgaaga ccactagatt
     6961 catcggaagc attgaagacc tacctgggga agtagcgaga cacctccagg ggtataacag
     7021 gtggatcacc cttgaggaca tcaggtccag atcatctctg ctagactaca gttgcttatg
     7081 acccggggga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     7141 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     7201 tgaaaggagg aactatatcc ggatccacta gttctagagc ggccgccacc gcggtggagc
     7261 tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     7321 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     7381 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     7441 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     7501 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     7561 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     7621 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     7681 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     7741 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     7801 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     7861 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     7921 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     7981 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     8041 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     8101 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     8161 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     8221 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     8281 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     8341 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     8401 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     8461 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     8521 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     8581 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     8641 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     8701 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     8761 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     8821 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     8881 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     8941 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     9001 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     9061 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     9121 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     9181 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     9241 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     9301 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     9361 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     9421 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctg
//
