LOCUS       sequence-53663-e        7931 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 7931)
  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 7931)
  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_J72110-BBa_J72103
FEATURES             Location/Qualifiers
     source          1..7931
                     /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    complement(4843..4867)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     rep_origin      4947..5492
                     /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     5393..5412
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             5915..6574
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5981..6000)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      6595..6689
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     protein_bind    complement(6702..6735)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        6773..6877
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6878..6946,6947..7738)
                     /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"
     primer_bind     complement(7096..7115)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    7838..7862
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      complement(7886..7929)
                     /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 tcctaactcg acgtgcaggc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4801 cggcgagcgg tatcagctca ctcaaaggcg gtaatcaatt cgacccagct ttcttgtaca
     4861 aagtggttga tccttacaga tcccggttat ccacagaatc aggggaggcc ttagaaatat
     4921 tttatctgat taataagatg atcttcttga gatcgttttg gtctgcgcgt aatctcttgc
     4981 tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga aggttctctg agctaccaac
     5041 tctttgaacc gaggtaactg gcttggagga gcgcagtcac caaaacttgt cctttcagtt
     5101 tagccttaac cggcgcatga cttcaagact aactcctcta aatcaattac cagtggctgc
     5161 tgccagtggt gcttttgcat gtctttccgg gttggactca agacgatagt taccggataa
     5221 ggcgcagcgg tcggactgaa cggggggttc gtgcatacag tccagcttgg agcgaactgc
     5281 ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg cggccataac agcggaatga
     5341 caccggtaaa ccgaaaggca ggaacaggag agcgcacgag ggagccgcca gggggaaacg
     5401 cctggtatct ttatagtcct gtcgggtttc gccaccactg atttgagcgt cagatttcgt
     5461 gatgcttgtc aggggggcgg agcctatgga aaaacggctt tgccgcggcc ctctcacttc
     5521 cctgttaagt atcttcctgg catcttccag gaaatctccg ccccgttcgt aagccatttc
     5581 cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag cggaatatat
     5641 cctgtatcac atattctgct gacgcaccgg tgcagccttt tttctcctgc cacatgaagc
     5701 acttcactga caccctcatc agtgccaaca tagtaagcca gtatacactc cgctagcgct
     5761 gatgtccggc ggtgcgcatg ccgttaaggg attttggtca tgaactagct tgatcgggca
     5821 cgtaagaggt tccaactttc accataatga aataagatca ctaccgggcg tattttttga
     5881 gttatcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac
     5941 caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc
     6001 tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa
     6061 gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc
     6121 tcatccggaa tttcgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca
     6181 cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata
     6241 ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga
     6301 aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc
     6361 ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc
     6421 cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat
     6481 tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca
     6541 acagtactgc gatgagtggc agggcggggc gtaatttgat atcgagctcg cttggactcc
     6601 tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga acgctcggtt
     6661 gccgccgggc gttttttatt ggtgagaatc caagcctgca gataacttcg tatagcatac
     6721 attatacgaa gttatctcga ggacgtcagg tggcactttt cggggaaatg tgcgcggaac
     6781 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6841 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6901 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6961 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     7021 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     7081 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     7141 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     7201 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     7261 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     7321 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     7381 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     7441 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     7501 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     7561 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     7621 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     7681 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     7741 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7801 aaggatctag gtgaagatcc ttgaattgcc attatcaaca agtttgtaca aaaaagcagg
     7861 ctccgaattg gtatcacgag gcagaatttc agataaaaaa aatccttagc tttcgctaag
     7921 gatgatttct g
//
