LOCUS       sequence-53658-e        4815 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 4815)
  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 4815)
  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_J72094
FEATURES             Location/Qualifiers
     source          1..4815
                     /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(1727..1751)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     rep_origin      1831..2376
                     /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     2277..2296
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2799..3458
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2865..2884)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3479..3573
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     protein_bind    complement(3586..3619)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3657..3761
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3762..3830,3831..4622)
                     /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(3980..3999)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    4722..4746
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      complement(4770..4813)
                     /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 aggatcctaa ctcgacgtgc aggcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1681 gctgcggcga gcggtatcag ctcactcaaa ggcggtaatc aattcgaccc agctttcttg
     1741 tacaaagtgg ttgatcctta cagatcccgg ttatccacag aatcagggga ggccttagaa
     1801 atattttatc tgattaataa gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc
     1861 ttgctctgaa aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc tctgagctac
     1921 caactctttg aaccgaggta actggcttgg aggagcgcag tcaccaaaac ttgtcctttc
     1981 agtttagcct taaccggcgc atgacttcaa gactaactcc tctaaatcaa ttaccagtgg
     2041 ctgctgccag tggtgctttt gcatgtcttt ccgggttgga ctcaagacga tagttaccgg
     2101 ataaggcgca gcggtcggac tgaacggggg gttcgtgcat acagtccagc ttggagcgaa
     2161 ctgcctaccc ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca taacagcgga
     2221 atgacaccgg taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc gccaggggga
     2281 aacgcctggt atctttatag tcctgtcggg tttcgccacc actgatttga gcgtcagatt
     2341 tcgtgatgct tgtcaggggg gcggagccta tggaaaaacg gctttgccgc ggccctctca
     2401 cttccctgtt aagtatcttc ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca
     2461 tttccgctcg ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag gaagcggaat
     2521 atatcctgta tcacatattc tgctgacgca ccggtgcagc cttttttctc ctgccacatg
     2581 aagcacttca ctgacaccct catcagtgcc aacatagtaa gccagtatac actccgctag
     2641 cgctgatgtc cggcggtgcg catgccgtta agggattttg gtcatgaact agcttgatcg
     2701 ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt
     2761 ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     2821 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     2881 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     2941 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     3001 atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg
     3061 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     3121 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     3181 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca
     3241 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     3301 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
     3361 cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat
     3421 tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tgatatcgag ctcgcttgga
     3481 ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc
     3541 ggttgccgcc gggcgttttt tattggtgag aatccaagcc tgcagataac ttcgtatagc
     3601 atacattata cgaagttatc tcgaggacgt caggtggcac ttttcgggga aatgtgcgcg
     3661 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3721 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3781 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3841 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3901 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3961 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4021 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4081 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4141 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4201 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4261 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4321 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4381 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4441 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4501 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4561 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4621 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4681 ttaaaaggat ctaggtgaag atccttgaat tgccattatc aacaagtttg tacaaaaaag
     4741 caggctccga attggtatca cgaggcagaa tttcagataa aaaaaatcct tagctttcgc
     4801 taaggatgat ttctg
//
