LOCUS       sequence-53667-e        7724 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7724)
  AUTHORS   Kittleson JT, DeLoache W, Cheng HY, Anderson JC
  TITLE     Scalable plasmid transfer using engineered P1-based phagemids.
  JOURNAL   ACS Synth Biol. 2012 Dec 21;1(12):583-9. doi: 10.1021/sb300054p. 
            Epub 2012 Aug 30.
  PUBMED    23656280
REFERENCE   2  (bases 1 to 7724)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1021/sb300054p";
            journalName: "ACS Synth Biol"; date: "2012-12-21- 21"; volume: "1"; 
            issue: "12"; pages: "583-9"
COMMENT     Sequence Label: BBa_J72113-BBa_J72152
FEATURES             Location/Qualifiers
     source          1..7724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(60..938)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        965..1249
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1172..1191
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      1501..1572
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1588..1615
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    4744..4762
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4769..4787
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             4816..4827
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4834..5547
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      5565..5636
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5652..5679
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    complement(5791..5815)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     rep_origin      5895..6440
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     6341..6360
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(6736..6830)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6851..7510)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7425..7444
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    7631..7655
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      complement(7679..7722)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 gaattcatga gatctctatg ctactccatc gagccgtcaa ttgtctgatt cgttaccaat
       61 tatgacaact tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc
      121 gggctggccc cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca
      181 acattgcgac cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg
      241 ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga
      301 tgtgacagac gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg
      361 ctgtctgcca ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt
      421 ggatggagcg actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt
      481 atcgccagca gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac
      541 aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat
      601 attgacggcc agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg
      661 tgataccatt cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac
      721 agcaaaatat cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga
      781 ccgcgaatgg tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa
      841 tcgagataac cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag
      901 tatcccggca gcaggggatc attttgcgct tcagccatac ttttcatact cccgccattc
      961 agagaagaaa ccaattgtcc atattgcatc agacattgcc gtcactgcgt cttttactgg
     1021 ctcttctcgc taaccaaacc ggtaaccccg cttattaaaa gcattctgta acaaagcggg
     1081 accaaagcca tgacaaaaac gcgtaacaaa agtgtctata atcacggcag aaaagtccac
     1141 attgattatt tgcacggcgt cacactttgc tatgccatag catttttatc cataagatta
     1201 gcggatctta cctgacgctt tttatcgcaa ctctctactg tttctccata cccggatctt
     1261 acagtgaggc ataattatgg ctttcattcc accaaccatc gacgacgtta gacattgctc
     1321 taacgcttta tctgtagacc ccgccgaaac cgacgctgcc cgcgccattg ctgaacacta
     1381 ctcaaagata tccaatcagg agtaccgcat cacccaagac gacctggatg atctcactga
     1441 cacaatcgaa tatctcatgg ccactaacca gccagactca caataaggat ctccaggcat
     1501 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     1561 gtgaacgctc tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat
     1621 aggatctcta ataggctatg tacgcgtatc aacaaatgaa caaaacactg ctttacaacg
     1681 aaacgctctt gaaagcgcag gatgtgagct aatttttgag gacaaggcga gcggcaaaaa
     1741 ggctgagcgc cctgggttaa aaaaggtact ccgtaacttt ccagaggtga taccctggtc
     1801 gtatggaagt tagaccgtct tgggcgcagc atgcgtcact tagttgtgct ggtggaagag
     1861 ctgcgtgaca gaggcattaa cttccggagt ctcactgact ccatcgacac cagtacacca
     1921 atggggcgct ttttctttca cgtaatgggg gcgctggcag aaatggaacg tgagcttatc
     1981 gttgaacgta cacgcgctgg acttgatgca gctcgcgcag aaggtcgtat aggtgggcgt
     2041 cggcctaaat accaagaaga aacatggcag caaatgcggc gattgctgga gaagggcatc
     2101 ccccgtaagc aggttgcaat catctatgat gtggctgttt ccacgcttta taagaagttt
     2161 ccggcgtcat catttcaatc ctaaaccttg gtttaagaga actcggggat cttaaagcga
     2221 ttttactttt tctggctatg aattggcctg ctttgtaaca cactccggtc tatcccgtag
     2281 cgccgggcat atcctgtcgc aatgtgcaaa tctcgcggca acaaccagtg aatacttcat
     2341 tcacaagcct caccgcctga tcgcggcaga aactggttat agccaatcaa ccgtcgttcg
     2401 tgcattccgt gaagctgtaa acaaaggtat tctgtctgta gagattgtta tcggcgatca
     2461 ccgtgaacgt cgcgctaacc tgtaccggtt tacaccatcc tttttggcct tcgcacaaca
     2521 agccaaaaat gcgctgatag aaagcaaatt aaagatttcg tcagcggcaa ccaaggttaa
     2581 agctgttctc gctaagacat tggctttatt taatttttta tccacacccc catgtcaaaa
     2641 tgataccccc tccccctgtc aggatgacgt ggcaataaag aataagaagt cacaagttaa
     2701 aaaaacaaaa agatcagttt ccggcggtgc cggaacaacc agcctcaaaa aattgacttc
     2761 atggatcgct aaggcaaaag caaaggctga caatctgcgg ttatccaaaa aacgcactca
     2821 aaaacatgag ttcaagcaga aagtagaggc ggctgcgcgg aaatatgctt acctgaaaaa
     2881 caagcgttcg cctgatattg gcgggatatc aaacttcgat aacctaccgc attgcatgac
     2941 ggtaaacgaa gctcttaatg cggttttagc caaaaataaa gataacgaac aatggggtat
     3001 accggcagga ttcagagggt aaggatctga atctggtgtg ttaaccgatt ctacgagtag
     3061 tcatttgttc attgagtagg aatattgttg ggatctcgaa aggaagcata agtgacctgg
     3121 gacgatcaca agaagaattt tgctcgcctg gcgcgagatg gtggttacac catcgcacag
     3181 tatgccgccg agtttaatct taaccctaat accgcacgtc gttatctccg tgccttcaaa
     3241 gaagacacca ggactacgga cagccgcaag ccaaataagc cagtcaggaa gccactaaaa
     3301 agcatgatca ttgatcactc taatgatcaa catgcaggtg atcacattgc ggctgaaata
     3361 gcggaaaaac aaagagttaa tgccgttgtc agtgccgcag tcgagaatgc gaagcgccaa
     3421 aataagcgca taaatgatcg ttcagatgat catgacgtga tcacccgcgc ccaccggacc
     3481 ttacgtgatc gcctggaacg cgacaccctg gatgatgatg gtgaacgctt tgagttcgaa
     3541 gttggcgatt acctgataga taacgttgaa gcgcggaagg ccgcgcgcgc tatgttgcgt
     3601 cggtccgggg ccgatgttct ggaaaccact cttctggaaa agtctctttc tcatctcctt
     3661 atgctggaga acgccaggga tacgtgtatt cgcctggtgc aggaaatgcg cgatcagcaa
     3721 aaagacgatg atgaaggtac tccgcctgaa taccgtatcg cgagcatgct aaacagctgt
     3781 tccgcgcaga taagcagcct gatcaacacc atttacagca tccggaataa ctatcgaaaa
     3841 gaaagccggg aggcggaaaa gcacgcttta tctatggggc aagctggcat tgttaagctg
     3901 gcatacgaac gaaagcgtga aaataactgg tcagtgctgg aagcggctga gttcatcgag
     3961 gcgcatggag gaaaagtgcc gcccctgatg ctggagcaaa tcaaagccga tctgcgtgct
     4021 cctaagacca ataccgatga tgaggaaaac caaacagcat ctggcgctcc atcacttgag
     4081 gatctggata aaatcgcgcg agaacgggcc gccagccgcc gcgctgatgc cgcattgtgg
     4141 attgagcatc gtagagaaga aattgccgat atcgtcgata caggtggtta tggtgatgtc
     4201 gatgcggaag gcatatcaaa cgaagcatgg cttgaacagg atctggacga agacgaggag
     4261 gaagacgaag aagttacccg caaactgtac ggggatgatg attaaggatc tgagcgattg
     4321 ctggcctaga tgaatgactg tccacgacag aacccggctt atcggtcagt ttcacctgat
     4381 ttacgtaaaa acccgcttcg gcgggttttt gcttttggag gggcggaaag atgaatgact
     4441 gtccacgacg ctatacccaa aagaaagcgg cttatcggtc agtttcacct ggtttacgta
     4501 aaaacccgct tcggcgggtt tttgcttttg gaggggcaga aagatgaatg actgtccacg
     4561 acactatacc caaaagaaag cggcttatcg gtcagtttca cctgttttac gtaaaaaccc
     4621 gcttcggcgg gtttttactt ttggaggggc agaaagatga atgactgtcc acgacactat
     4681 acccaaaaga aagcggctta tcggtcagtt ttacctgatg tacgtaataa accgttcgga
     4741 tcttccctat cagtgataga gattgacatc cctatcagtg atagagatac tgagcacgga
     4801 tcttagctac tagagaaaga ggagaaatac tagatgcgta aaggcgaaga gctgttcact
     4861 ggtgtcgtcc ctattctggt ggaactggat ggtgatgtca acggtcataa gttttccgtg
     4921 cgtggcgagg gtgaaggtga cgcaactaat ggtaaactga cgctgaagtt catctgtact
     4981 actggtaaac tgccggtacc ttggccgact ctggtaacga cgctgactta tggtgttcag
     5041 tgctttgctc gttatccgga ccatatgaag cagcatgact tcttcaagtc cgccatgccg
     5101 gaaggctatg tgcaggaacg cacgatttcc tttaaggatg acggcacgta caaaacgcgt
     5161 gcggaagtga aatttgaagg cgataccctg gtaaaccgca ttgagctgaa aggcattgac
     5221 tttaaagaag acggcaatat cctgggccat aagctggaat acaattttaa cagccacaat
     5281 gtttacatca ccgccgataa acaaaaaaat ggcattaaag cgaattttaa aattcgccac
     5341 aacgtggagg atggcagcgt gcagctggct gatcactacc agcaaaacac tccaatcggt
     5401 gatggtcctg ttctgctgcc agacaatcac tatctgagca cgcaaagcgt tctgtctaaa
     5461 gatccgaacg agaaacgcga tcatatggtt ctgctggagt tcgtaaccgc agcgggcatc
     5521 acgcatggta tggatgaact gtacaaatga ggatctccag gcatcaaata aaacgaaagg
     5581 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact
     5641 agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttataggatc ctaactcgac
     5701 gtgcaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5761 tcagctcact caaaggcggt aatcaattcg acccagcttt cttgtacaaa gtggttgatc
     5821 cttacagatc ccggttatcc acagaatcag gggaggcctt agaaatattt tatctgatta
     5881 ataagatgat cttcttgaga tcgttttggt ctgcgcgtaa tctcttgctc tgaaaacgaa
     5941 aaaaccgcct tgcagggcgg tttttcgaag gttctctgag ctaccaactc tttgaaccga
     6001 ggtaactggc ttggaggagc gcagtcacca aaacttgtcc tttcagttta gccttaaccg
     6061 gcgcatgact tcaagactaa ctcctctaaa tcaattacca gtggctgctg ccagtggtgc
     6121 ttttgcatgt ctttccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6181 ggactgaacg gggggttcgt gcatacagtc cagcttggag cgaactgcct acccggaact
     6241 gagtgtcagg cgtggaatga gacaaacgcg gccataacag cggaatgaca ccggtaaacc
     6301 gaaaggcagg aacaggagag cgcacgaggg agccgccagg gggaaacgcc tggtatcttt
     6361 atagtcctgt cgggtttcgc caccactgat ttgagcgtca gatttcgtga tgcttgtcag
     6421 gggggcggag cctatggaaa aacggctttg ccgcggccct ctcacttccc tgttaagtat
     6481 cttcctggca tcttccagga aatctccgcc ccgttcgtaa gccatttccg ctcgccgcag
     6541 tcgaacgacc gagcgtagcg agtcagtgag cgaggaagcg gaatatatcc tgtatcacat
     6601 attctgctga cgcaccggtg cagccttttt tctcctgcca catgaagcac ttcactgaca
     6661 ccctcatcag tgccaacata gtaagccagt atacactccg ctagcgctga tgtccggcgg
     6721 tgcctcgagg cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa
     6781 caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct
     6841 cgatatcaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca
     6901 ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca
     6961 gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt
     7021 ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga
     7081 aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca
     7141 catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg
     7201 atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata
     7261 tcaccagctc accgtctttc attgccatac gaaattccgg atgagcattc atcaggcggg
     7321 caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa
     7381 aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg
     7441 cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt
     7501 ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
     7561 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgcccg atcaagaatt
     7621 gccattatca acaagtttgt acaaaaaagc aggctccgaa ttggtatcac gaggcagaat
     7681 ttcagataaa aaaaatcctt agctttcgct aaggatgatt tctg
//
