LOCUS       sequence-60656-e        6614 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 6614)
  AUTHORS   Bonnet J, Yin P, Ortiz ME, Subsoontorn P, Endy D
  TITLE     Amplifying genetic logic gates.
  JOURNAL   Science. 2013 May 3;340(6132):599-603. doi: 10.1126/science.1232758.
            Epub 2013 Mar 28.
  PUBMED    23539178
REFERENCE   2  (bases 1 to 6614)
  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: 
            "10.1126/science.1232758"; journalName: "Science"; date: "2013-05-3-
            3"; volume: "340"; issue: "6132"; pages: "599-603"
COMMENT     Sequence Label: Dual-recombinase-controller
FEATURES             Location/Qualifiers
     source          1..6614
                     /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"""
     protein_bind    23..41
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    48..66
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             111..1610
                     /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"
     terminator      1639..1666
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1749..2627)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2654..2937
                     /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     2860..2879
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2991..3008
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4476..4508
                     /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="AAKDENYALAA"
     misc_feature    4512..4532
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(4616..4633)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4839..5427)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4919..4938)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(5515..5609)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5630..6289)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6204..6223
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(6290..6392)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6569..6612)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 gaattcgcgg ccgcttctag agtccctatc agtgatagag attgacatcc ctatcagtga
       61 tagagatact gagcacaagc ttgcggcttt cacacttggg ggctagcgtg agagccctgg
      121 tagtcatccg cctgtcccgc gtcaccgatg ctacgacttc accggagcgt cagctggagt
      181 cttgccagca gctctgcgcc cagcgcggct gggacgtcgt cggggtagcg gaggatctgg
      241 acgtctccgg ggcggtcgat ccgttcgacc ggaagcgcag accgaacctg gcccggtggc
      301 tagcgttcga ggagcaaccg tttgacgtga tcgtggcgta ccgggtagat cggttgaccc
      361 gatcgatccg gcatcttcag cagctggtcc actgggccga ggaccacaag aagctggtcg
      421 tctccgcgac cgaagcgcac ttcgatacga cgacgccgtt tgcggcggtc gtcatcgcgc
      481 ttatgggaac ggtggcgcag atggaattag aagcgatcaa agagcggaac cgttcggctg
      541 cgcatttcaa tatccgcgcc gggaaatacc gaggatccct gccgccgtgg ggatacctgc
      601 ctacgcgcgt ggacggggag tggcggctgg tgccggaccc tgtgcagcga gagcgcatcc
      661 tcgaggtgta tcaccgcgtc gtcgacaacc acgagccgct gcatctggtg gcccacgacc
      721 tgaaccggcg tggtgtcctg tcgccgaagg actacttcgc gcagctgcaa ggccgcgagc
      781 cgcagggccg ggagtggtcg gctaccgcgc tgaagcgatc gatgatctcc gaggcgatgc
      841 tcgggtacgc gactctgaac ggtaagaccg tccgagacga cgacggagcc ccgctggtgc
      901 gggctgagcc gatcctgacc cgtgagcagc tggaggcgct gcgcgccgag ctcgtgaaga
      961 cctcccgggc gaagcccgcg gtgtctaccc cgtcgctgct gctgcgggtg ttgttctgcg
     1021 cggtgtgcgg ggagcccgcg tacaagttcg ccgggggagg acgtaagcac ccgcgctacc
     1081 gctgccgctc gatggggttc ccgaagcact gcgggaacgg cacggtggcg atggccgagt
     1141 gggacgcgtt ctgcgaggag caggtactgg atctgctcgg ggacgcggag cgtctggaga
     1201 aagtctgggt agcgggctcg gactccgcgg tcgaactcgc ggaggtgaac gcggagctgg
     1261 tggacctgac gtcgctgatc ggctccccgg cctaccgggc gggctctccg cagcgagaag
     1321 cactggatgc ccgtattgcg gcgctggccg cgcggcaaga ggagctggag ggcctggagg
     1381 ctcgcccgtc tggctgggag tggcgcgaga ccgggcagcg gttcggggac tggtggcggg
     1441 agcaggacac cgcggcaaag aacacctggc ttcggtcgat gaacgttcgg ctgacgttcg
     1501 acgtccgcgg cgggctgact cgcacgatcg acttcgggga tcttcaggag tacgagcagc
     1561 atctcaggct cggcagcgtg gtcgaacggc tacacaccgg gatgtcgtaa taatactaga
     1621 tgcattaata atcacactgg ctcaccttcg ggtgggcctt tctgcgttta tatactagag
     1681 agagaatata aaaagccaga ttattaatcc ggctttttta ttatttcctg cagggcggcc
     1741 gccatatgtt atgacaactt gacggctaca tcattcactt tttcttcaca accggcacgg
     1801 aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaaata gagttgatcg
     1861 tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct caaaagcagc
     1921 ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc taactgctgg
     1981 cggaaaagat gtgacagacg cgacggcgac aagcaaacat gctgtgcgac gctggcgata
     2041 tcaaaattgc tgtctgccag gtgatcgctg atgtactgac aagcctcgcg tacccgatta
     2101 tccatcggtg gatggagcga ctcgttaatc gcttccatgc gccgcagtaa caattgctca
     2161 agcagattta tcgccagcag ctccgaatag cgcccttccc cttgcccggc gttaatgatt
     2221 tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta
     2281 ttggcaaata ttgacggcca gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa
     2341 acccactggt gataccattc gcgagcctcc ggatgacgac cgtagtgatg aatctctcct
     2401 ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg atttttcacc
     2461 accccctgac cgcgaatggt gagattgaga atataacctt tcattcccag cggtcggtcg
     2521 ataaaaaaat cgagataacc gttggcctca atcggcgtta aacccgccac cagatgggca
     2581 ttaaacgagt atcccggcag caggggatca ttttgcgctt cagccatact tttcatactc
     2641 ccgccattca gagaagaaac caattgtcca tattgcatca gacattgccg tcactgcgtc
     2701 ttttactggc tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa
     2761 caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga
     2821 aaagtcacat tgattatttg cacggcgtca cactttgcta tgccatagca tttttatcca
     2881 taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt tctccatacc
     2941 gtttttttgg gctagctact agagtcacac aggaaaccta ctagatgaaa catcatcacc
     3001 atcaccacca ggccggcact aagaaagtag caatctatac acgagtatcc actactaacc
     3061 aagcagagga aggcttctca attgatgagc aaattgaccg tttaacaaaa tatgctgaag
     3121 caatggggtg gcaagtatct gatacttata ctgatgctgg tttttcaggg gccaaacttg
     3181 aacgcccagc aatgcaaaga ttaatcaacg atatcgagaa taaagctttt gatacagttc
     3241 ttgtatataa gctagaccgc ctttcacgta gtgtaagaga tactctttat cttgttaagg
     3301 atgtgttcac aaaaaataaa atagacttta tctcgcttaa tgaaagtatt gatacttctt
     3361 ctgctatggg tagcttgttt ctcactattc tttctgcaat taatgagttt gaaagagaga
     3421 atataaaaga acgcatgact atgggtaaac tagggcgagc gaaatctggt aagtctatga
     3481 tgtggactaa gacagctttt gggtattacc acaacagaaa gacaggtata ttagaaattg
     3541 ttcctttaca agctacaata gttgaacaaa tattcactga ttatttatca ggaatatcac
     3601 ttacaaaatt aagagataaa ctcaatgaat ctggacacat cggtaaagat ataccgtggt
     3661 cttatcgtac cctaagacaa acacttgata atccagttta ctgtggttat atcaaattta
     3721 aggacagcct atttgaaggt atgcacaaac caattatccc ttatgagact tatttaaaag
     3781 ttcaaaaaga gctagaagaa agacaacagc agacttatga aagaaataac aaccctagac
     3841 ctttccaagc taaatatatg ctgtcaggga tggcaaggtg cggttactgt ggagcacctt
     3901 taaaaattgt tcttggccac aaaagaaaag atggaagccg cactatgaaa tatcactgtg
     3961 caaatagatt tcctcgaaaa acaaaaggaa ttacagtata taatgacaat aaaaagtgtg
     4021 attcaggaac ttatgattta agtaatttag aaaatactgt tattgacaac ctgattggat
     4081 ttcaagaaaa taatgactcc ttattgaaaa ttatcaatgg caacaaccaa cctattcttg
     4141 atacttcgtc atttaaaaag caaatttcac agatcgataa aaaaatacaa aagaactctg
     4201 atttgtacct aaatgatttt atcactatgg atgagttgaa agatcgtact gattcccttc
     4261 aggctgagaa aaagctgctt aaagctaaga ttagcgaaaa taaatttaat gactctactg
     4321 atgtttttga gttagttaaa actcagttgg gctcaattcc gattaatgaa ctatcatatg
     4381 ataataaaaa gaaaatcgtc aacaaccttg tatcaaaggt tgatgttact gctgataatg
     4441 tagatatcat atttaaattc caactcgcta ccggtgctgc taaggacgaa aactacgctc
     4501 tggctgctta atactagtag cggccgctgc agtccggcaa aaaaacgggc aaggtgtcac
     4561 caccctgccc tttttcttta aaaccgaaaa gattacttcg cgttatgcag gcttcctcgc
     4621 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4681 cggtaatcct agggcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4741 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4801 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4861 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4921 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4981 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     5041 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5101 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5161 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5221 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5281 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5341 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5401 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5461 gctcagtgga acgaaaactc acgttaaggg attttggtca tgacttgtgc ttggattctc
     5521 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga
     5581 ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat tacgccccgc
     5641 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
     5701 cacagacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
     5761 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
     5821 caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc
     5881 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta
     5941 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct
     6001 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
     6061 ttgccatacg aaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
     6121 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
     6181 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
     6241 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
     6301 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat
     6361 ggtgaaagtt ggaacctctt acgtgccgat caacgtctca ttttcgccag atatcgacgt
     6421 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     6481 tcgtcttcac ctcgagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     6541 tataaaaata ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt agctttcgct
     6601 aaggatgatt tctg
//
