LOCUS       sequence-48285-e        5881 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 5881)
  AUTHORS   Bonnet J, Subsoontorn P, Endy D
  TITLE     Rewritable digital data storage in live cells via engineered control
            of recombination directionality.
  JOURNAL   Proc Natl Acad Sci U S A. 2012 Jun 5;109(23):8884-9. Epub 2012 May 
            21.
  PUBMED    22615351
REFERENCE   2  (bases 1 to 5881)
  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"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2012-06-5"; volume: "109(23)"; pages: 
            "8884-9. Epub 2012 May 21"
COMMENT     Sequence Label: pBad-Int Set Flipper
FEATURES             Location/Qualifiers
     source          1..5881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..22
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(23..901)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        928..1211
                     /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     1134..1153
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1265..1282
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1292..2788
                     /codon_start=1
                     /transl_table=11
                     /gene="Bxb1"
                     /product="Bxb1 integrase"
                     /label=Bxb1 integrase
                     /note="Bxb1 large serine integrase from Mycobacterium 
                     bacteriophage Bxb1 (Ghosh et al. 2003)."
                     /protein_id="AFF61396.1"
                     /translation="RALVVIRLSRVTDATTSPERQLESCQQLCAQRGWDVVGVAEDLDV
                     SGAVDPFDRKRRPNLARWLAFEEQPFDVIVAYRVDRLTRSIRHLQQLVHWAEDHKKLVV
                     SATEAHFDTTTPFAAVVIALMGTVAQMELEAIKERNRSAAHFNIRAGKYRGSLPPWGYL
                     PTRVDGEWRLVPDPVQRERILEVYHRVVDNHEPLHLVAHDLNRRGVLSPKDYFAQLQGR
                     EPQGREWSATALKRSMISEAMLGYATLNGKTVRDDDGAPLVRAEPILTREQLEALRAEL
                     VKTSRAKPAVSTPSLLLRVLFCAVCGEPAYKFAGGGRKHPRYRCRSMGFPKHCGNGTVA
                     MAEWDAFCEEQVLDLLGDAERLEKVWVAGSDSAVELAEVNAELVDLTSLIGSPAYRAGS
                     PQREALDARIAALAARQEELEGLEARPSGWEWRETGQRFGDWWREQDTAAKNTWLRSMN
                     VRLTFDVRGGLTRTIDFGDLQEYEQHLRLGSVVERLHTGMS"
     CDS             2795..2827
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag
                     /note="wild-type LAA variant"
                     /translation="AANDENYALAA"
     misc_feature    2840..2860
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     rep_origin      3339..3884
                     /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     3785..3804
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4001..4018
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4468..4487)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4479..5294
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4479..5294
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4552..4571)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gaattcgcgg ccgcttctag agttatgaca acttgacggc tacatcattc actttttctt
       61 cacaaccggc acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga
      121 aatagagttg atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg
      181 tgctcaaaag cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa
      241 tccctaactg ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg
      301 cgacgctggc gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct
      361 cgcgtacccg attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca
      421 gtaacaattg ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc
      481 cggcgttaat gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc
      541 gaaagaaccc cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc
      601 gcggacgaaa gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt
      661 gatgaatctc tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc
      721 cctgattttt caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc
      781 ccagcggtcg gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg
      841 ccaccagatg ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca
      901 tacttttcat actcccgcca ttcagagaag aaaccaattg tccatattgc atcagacatt
      961 gccgtcactg cgtcttttac tggctcttct cgctaaccaa accggtaacc ccgcttatta
     1021 aaagcattct gtaacaaagc gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct
     1081 ataatcacgg cagaaaagtc acattgatta tttgcacggc gtcacacttt gctatgccat
     1141 agcattttta tccataagat tagcggatcc tacctgacgc tttttatcgc aactctctac
     1201 tgtttctcca taccgttttt ttgggctagc tactagagtc acacaggaaa cctactagat
     1261 gaaacatcat caccatcacc accaggccgg cagagccctg gtagtcatcc gcctgtcccg
     1321 cgtcaccgat gctacgactt caccggagcg tcagctggag tcttgccagc agctctgcgc
     1381 ccagcgcggc tgggacgtcg tcggggtagc ggaggatctg gacgtctccg gggcggtcga
     1441 tccgttcgac cggaagcgca gaccgaacct ggcccggtgg ctagcgttcg aggagcaacc
     1501 gtttgacgtg atcgtggcgt accgggtaga tcggttgacc cgatcgatcc ggcatcttca
     1561 gcagctggtc cactgggccg aggaccacaa gaagctggtc gtctccgcga ccgaagcgca
     1621 cttcgatacg acgacgccgt ttgcggcggt cgtcatcgcg cttatgggaa cggtggcgca
     1681 gatggaatta gaagcgatca aagagcggaa ccgttcggct gcgcatttca atatccgcgc
     1741 cgggaaatac cgaggatccc tgccgccgtg gggatacctg cctacgcgcg tggacgggga
     1801 gtggcggctg gtgccggacc ctgtgcagcg agagcgcatc ctcgaggtgt atcaccgcgt
     1861 cgtcgacaac cacgagccgc tgcatctggt ggcccacgac ctgaaccggc gtggtgtcct
     1921 gtcgccgaag gactacttcg cgcagctgca aggccgcgag ccgcagggcc gggagtggtc
     1981 ggctaccgcg ctgaagcgat cgatgatctc cgaggcgatg ctcgggtacg cgactctgaa
     2041 cggtaagacc gtccgagacg acgacggagc cccgctggtg cgggctgagc cgatcctgac
     2101 ccgtgagcag ctggaggcgc tgcgcgccga gctcgtgaag acctcccggg cgaagcccgc
     2161 ggtgtctacc ccgtcgctgc tgctgcgggt gttgttctgc gcggtgtgcg gggagcccgc
     2221 gtacaagttc gccgggggag gacgtaagca cccgcgctac cgctgccgct cgatggggtt
     2281 cccgaagcac tgcgggaacg gcacggtggc gatggccgag tgggacgcgt tctgcgagga
     2341 gcaggtactg gatctgctcg gggacgcgga gcgtctggag aaagtctggg tagcgggctc
     2401 ggactccgcg gtcgaactcg cggaggtgaa cgcggagctg gtggacctga cgtcgctgat
     2461 cggctccccg gcctaccggg cgggctctcc gcagcgagaa gcactggatg cccgtattgc
     2521 ggcgctggcc gcgcggcaag aggagctgga gggcctggag gctcgcccgt ctggctggga
     2581 gtggcgcgag accgggcagc ggttcgggga ctggtggcgg gagcaggaca ccgcggcaaa
     2641 gaacacctgg cttcggtcga tgaacgttcg gctgacgttc gacgtccgcg gcgggctgac
     2701 tcgcacgatc gacttcgggg atcttcagga gtacgagcag catctcaggc tcggcagcgt
     2761 ggtcgaacgg ctacacaccg ggatgtcgac cggcgctgct aacgacgaaa actacgctct
     2821 ggctgcttaa accggttaat actagtagcg gccgctgcag caatggcaac aacgttgcgc
     2881 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2941 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3001 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     3061 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3121 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     3181 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3241 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3301 ttccactgag cgtcagaccc cttaataaga tgatcttctt gagatcgttt tggtctgcgc
     3361 gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc
     3421 tgagctacca actctttgaa ccgaggtaac tggcttggag gagcgcagtc accaaaactt
     3481 gtcctttcag tttagcctta accggcgcat gacttcaaga ctaactcctc taaatcaatt
     3541 accagtggct gctgccagtg gtgcttttgc atgtctttcc gggttggact caagacgata
     3601 gttaccggat aaggcgcagc ggtcggactg aacggggggt tcgtgcatac agtccagctt
     3661 ggagcgaact gcctacccgg aactgagtgt caggcgtgga atgagacaaa cgcggccata
     3721 acagcggaat gacaccggta aaccgaaagg caggaacagg agagcgcacg agggagccgc
     3781 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc
     3841 gtcagatttc gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg
     3901 ccctctcact tccctgttaa gtatcttcct ggcatcttcc aggaaatctc cgccccgttc
     3961 gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga
     4021 agcggaatat atcctgtatc acatattctg ctgacgcacc ggtgcagcct tttttctcct
     4081 gccacatgaa gcacttcact gacaccctca tcagtgccaa catagtaagc cagtatacac
     4141 tccgctagcg ctgaggtctg cctcgtgaag aaggtgttgc tgactcatac caggcctgaa
     4201 tcgccccatc atccagccag aaagtgaggg agccacggtt gatgagagct ttgttgtagg
     4261 tggaccagtt ggtgattttg aacttttgct ttgccacgga acggtctgcg ttgtcgggaa
     4321 gatgcgtgat ctgatccttc aactcagcaa aagttcgatt tattcaacaa agccacgttg
     4381 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
     4441 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     4501 cgtcttgctc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     4561 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg
     4621 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     4681 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     4741 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc
     4801 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     4861 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     4921 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     4981 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat
     5041 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     5101 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     5161 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     5221 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     5281 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga
     5341 cttgacggga cggcggcttt gttgaataaa tcgaactttt gctgagttga aggatcagat
     5401 cacgcatctt cccgacaacg cagaccgttc cgtggcaaag caaaagttca aaatcaccaa
     5461 ctggtccacc tacaacaaag ctctcatcaa ccgtggctcc ctcactttct ggctggatga
     5521 tggggcgatt caggcctggt atgagtcagc aacaccttct tcacgaggca gacctcagcg
     5581 ctcaaagatg caggggtaaa agctaaccgc atctttaccg acaaggcatc cggcagttca
     5641 acagatcggg aagggctgga tttgctgagg atgaaggtgg aggaaggtga tgtcattctg
     5701 gtgaagaagc tcgaccgtct tggccgcgac accgccgaca tgatccaact gataaaagag
     5761 tttgatgctc agggtgtagc ggttcggttt attgacgacg ggatcagtac cgacggtgat
     5821 atggggcaaa tggtggtcac catcctgtcg gctgtggcac aggctgaacg ccggaggatc
     5881 c
//
