LOCUS       sequence-177674-        6375 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Synthetic gene circuit .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6375)
  AUTHORS   Rubens JR, Selvaggio G, Lu TK
  TITLE     Synthetic mixed-signal computation in living cells.
  JOURNAL   Nat Commun. 2016 Jun 3;7:11658. doi: 10.1038/ncomms11658.
  PUBMED    27255669
REFERENCE   2  (bases 1 to 6375)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2016-06-3"; volume: "7:11658. doi"; pages: " 
            10.1038/ncomms11658"
COMMENT     Sequence Label: pZS2oxySp*-RBS30-Bxbi-proD-oxyR
FEATURES             Location/Qualifiers
     source          1..6375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1125..1211
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1756..2706)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2754..2976
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3474..3568)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3599..4393)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3710..3729)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4320..4339
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     RBS             4646..4657
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4664..6163
                     /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="MRALVVIRLSRVTDATTSPERQLESCQQLCAQRGWDVVGVAEDLD
                     VSGAVDPFDRKRRPNLARWLAFEEQPFDVIVAYRVDRLTRSIRHLQQLVHWAEDHKKLV
                     VSATEAHFDTTTPFAAVVIALMGTVAQMELEAIKERNRSAAHFNIRAGKYRGSLPPWGY
                     LPTRVDGEWRLVPDPVQRERILEVYHRVVDNHEPLHLVAHDLNRRGVLSPKDYFAQLQG
                     REPQGREWSATALKRSMISEAMLGYATLNGKTVRDDDGAPLVRAEPILTREQLEALRAE
                     LVKTSRAKPAVSTPSLLLRVLFCAVCGEPAYKFAGGGRKHPRYRCRSMGFPKHCGNGTV
                     AMAEWDAFCEEQVLDLLGDAERLEKVWVAGSDSAVELAEVNAELVDLTSLIGSPAYRAG
                     SPQREALDARIAALAARQEELEGLEARPSGWEWRETGQRFGDWWREQDTAAKNTWLRSM
                     NVRLTFDVRGGLTRTIDFGDLQEYEQHLRLGSVVERLHTGMS"
     terminator      6271..6357
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cacagctaac accacgtcgt ccctatctgc tgccctaggt ctatgagtgg ttgctggata
       61 actttacggg catgcataag gctcgtataa tatattcagg gagaccacaa cggtttccct
      121 ctacaaataa ttttgtttaa ctttgaattc ttcacacagg aaaccggtac catgaatatt
      181 cgtgatcttg agtacctggt ggcattggct gaacaccgcc attttcggcg tgcggcagat
      241 tcctgccacg ttagccagcc gacgcttagc gggcaaattc gtaagctgga agatgagctg
      301 ggcgtgatgt tgctggagcg gaccagccgt aaagtgttgt tcacccaggc gggaatgctg
      361 ctggtggatc aggcgcgtac cgtgctgcgt gaggtgaaag tccttaaaga gatggcaagc
      421 cagcagggcg agacgatgtc cggaccgctg cacattggtt tgattcccac agttggaccg
      481 tacctgctac cgcatattat ccctatgctg caccagacct ttccaaagct ggaaatgtat
      541 ctgcatgaag cacagaccca ccagttactg gcgcaactgg acagcggcaa actcgattgc
      601 gtgatcctcg cgctggtgaa agagagcgaa gcattcattg aagtgccgtt gtttgatgag
      661 ccaatgttgc tggctatcta tgaagatcac ccgtgggcga accgcgaatg cgtaccgatg
      721 gccgatctgg caggggaaaa actgctgatg ctggaagatg gtcactgttt gcgcgatcag
      781 gcaatgggtt tctgttttga agccggggcg gatgaagata cacacttccg cgcgaccagc
      841 ctggaaactc tgcgcaacat ggtggcggca ggtagcggga tcactttact gccagcgctg
      901 gctgtgccgc cggagcgcaa acgcgatggg gttgtttatc tgccgtgcat taagccggaa
      961 ccacgccgca ctattggcct ggtttatcgt cctggctcac cgctgcgcag ccgctatgag
     1021 cagctggcag aggccatccg cgcaagaatg gatggccatt tcgataaagt tttaaaacag
     1081 gcggtttaac ccgggggatc ccatggtacg cgtgctagag gcatcaaata aaacgaaagg
     1141 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     1201 gtaggacaaa tccgccgccc tagacctagg gcctagggta cgggttttgc tgcccgcaaa
     1261 cgggctgttc tggtgttgct agtttgttat cagaatcgca gatccggctt cagccggttt
     1321 gccggctgaa agcgctattt cttccagaat tgccatgatt ttttccccac gggaggcgtc
     1381 actggctccc gtgttgtcgg cagctttgat tcgataagca gcatcgcctg tttcaggctg
     1441 tctatgtgtg actgttgagc tgtaacaagt tgtctcaggt gttcaatttc atgttctagt
     1501 tgctttgttt tactggtttc acctgttcta ttaggtgtta catgctgttc atctgttaca
     1561 ttgtcgatct gttcatggtg aacagctttg aatgcaccaa aaactcgtaa aagctctgat
     1621 gtatctatct tttttacacc gttttcatct gtgcatatgg acagttttcc ctttgatatg
     1681 taacggtgaa cagttgttct acttttgttt gttagtcttg atgcttcact gatagataca
     1741 agagccataa gaacctcaga tccttccgta tttagccagt atgttctcta gtgtggttcg
     1801 ttgtttttgc gtgagccatg agaacgaacc attgagatca tacttacttt gcatgtcact
     1861 caaaaatttt gcctcaaaac tggtgagctg aatttttgca gttaaagcat cgtgtagtgt
     1921 ttttcttagt ccgttatgta ggtaggaatc tgatgtaatg gttgttggta ttttgtcacc
     1981 attcattttt atctggttgt tctcaagttc ggttacgaga tccatttgtc tatctagttc
     2041 aacttggaaa atcaacgtat cagtcgggcg gcctcgctta tcaaccacca atttcatatt
     2101 gctgtaagtg tttaaatctt tacttattgg tttcaaaacc cattggttaa gccttttaaa
     2161 ctcatggtag ttattttcaa gcattaacat gaacttaaat tcatcaaggc taatctctat
     2221 atttgccttg tgagttttct tttgtgttag ttcttttaat aaccactcat aaatcctcat
     2281 agagtatttg ttttcaaaag acttaacatg ttccagatta tattttatga atttttttaa
     2341 ctggaaaaga taaggcaata tctcttcact aaaaactaat tctaattttt cgcttgagaa
     2401 cttggcatag tttgtccact ggaaaatctc aaagccttta accaaaggat tcctgatttc
     2461 cacagttctc gtcatcagct ctctggttgc tttagctaat acaccataag cattttccct
     2521 actgatgttc atcatctgag cgtattggtt ataagtgaac gataccgtcc gttctttcct
     2581 tgtagggttt tcaatcgtgg ggttgagtag tgccacacag cataaaatta gcttggtttc
     2641 atgctccgtt aagtcatagc gactaatcgc tagttcattt gctttgaaaa caactaattc
     2701 agacatacat ctcaattggt ctaggtgatt ttaatcacta taccaattga gatgggctag
     2761 tcaatgataa ttactagtcc ttttcctttg agttgtgggt atctgtaaat tctgctagac
     2821 ctttgctgga aaacttgtaa attctgctag accctctgta aattccgcta gacctttgtg
     2881 tgtttttttt gtttatattc aagtggttat aatttataga ataaagaaag aataaaaaaa
     2941 gataaaaaga atagatccca gccctgtgta taactcacta ctttagtcag ttccgcagta
     3001 ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt aaaaccctaa
     3061 aggcttaagt agcaccctcg caagctcggg caaatcgctg aatattcctt ttgtctccga
     3121 ccatcaggca cctgagtcgc tgtctttttc gtgacattca gttcgctgcg ctcacggctc
     3181 tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt atggattcat gcaaggaaac
     3241 tacccataat acaagaaaag cccgtcacgg gcttctcagg gcgttttatg gcgggtctgc
     3301 tatgtggtgc tatctgactt tttgctgttc agcagttcct gccctctgat tttccagtct
     3361 gaccacttcg gattatcccg tgacaggtca ttcagactgg ctaatgcacc cagtaaggca
     3421 gcggtatcat caacaggctt acccgtctta ctgtccctag tactagtgct tggattctca
     3481 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag
     3541 gtcattactg gatctatcaa caggagtcca agcgagctct cgaaccccag agtcccgctc
     3601 agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac
     3661 cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg
     3721 tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc
     3781 cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga
     3841 cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg gctggcgcga
     3901 gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac
     3961 gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg
     4021 tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag
     4081 atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag
     4141 tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg
     4201 ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg
     4261 ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg
     4321 cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat
     4381 cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagatctt gatcccctgc
     4441 gccatcagat ccttggcggc aagaaagcca tccagtttac tttgcagggc ttcccaacct
     4501 taccagaggg cgccccagct ggcaattccg acgtcttcat tatccatcct ccatcgccac
     4561 gatagttcat ggcgataggt agaatagcaa tgaacgatta tccctatcaa gcattctgac
     4621 tgagcattgc tcacacgaat tcattaaaga ggagaaaggt accatgagag ccctggtagt
     4681 catccgcctg tcccgcgtca ccgatgctac gacttcaccg gagcgtcagc tggagtcttg
     4741 ccagcagctc tgcgcccagc gcggctggga cgtcgtcggg gtagcggagg atctggacgt
     4801 ctccggggcg gtcgatccgt tcgaccggaa gcgcagaccg aacctggccc ggtggctagc
     4861 gttcgaggag caaccgttcg acgtgatcgt ggcgtaccgg gtagaccggt tgacccgatc
     4921 gatccggcat ctgcagcagc tggtccactg ggccgaggac cacaagaagc tggtcgtctc
     4981 cgcgaccgaa gcgcacttcg atacgacgac gccgtttgcg gcggtcgtca tcgcgcttat
     5041 gggaacggtg gcgcagatgg aattagaagc gatcaaagag cggaaccgtt cggctgcgca
     5101 tttcaatatc cgcgccggga aataccgagg atccctgccg ccgtggggat acctgcctac
     5161 gcgcgtggac ggggagtggc ggctggtgcc ggaccctgtg cagcgagagc gcatcctcga
     5221 ggtgtatcac cgcgtcgtcg acaaccacga gccgctgcac ctggtggccc acgacctgaa
     5281 ccggcgtggt gtcctgtcgc cgaaggacta cttcgcgcag ctgcaaggcc gcgagccgca
     5341 gggccgggag tggtcggcta ccgcgctgaa gcgatcgatg atctccgagg cgatgctcgg
     5401 gtacgcgact ctgaacggta agaccgtccg agacgacgac ggagccccgc tggtgcgggc
     5461 tgagccgatc ctgacccgtg agcagctgga ggcgctgcgc gccgagctcg tgaagacctc
     5521 ccgggcgaag cccgcggtgt ctaccccgtc gctgctgctg cgggtgttgt tctgtgcggt
     5581 gtgcggggag cccgcgtaca agttcgccgg gggaggacgt aagcacccgc gctaccgctg
     5641 ccgctcgatg gggttcccga agcactgcgg gaacggcacg gtggcgatgg ccgagtggga
     5701 cgcgttctgc gaggagcagg tgctggatct gctcggggac gcggagcgtc tggagaaagt
     5761 ctgggtagcc ggctcggact ccgcggtcga actcgcggag gtgaacgcgg agctggtgga
     5821 cctgacgtcg ctgatcggct ccccggccta ccgggccggc tctccgcagc gagaagcact
     5881 ggatgcccgt attgcggcgc tggccgcgcg gcaagaggag ctggagggtc tagaggctcg
     5941 cccgtctggc tgggagtggc gcgagaccgg gcagcggttc ggggactggt ggcgggagca
     6001 ggacaccgcg gcaaagaaca cctggcttcg gtcgatgaac gttcggctga cgttcgacgt
     6061 ccgcggcggg ctgactcgca cgatcgactt cggggatctg caggagtacg agcagcatct
     6121 caggctcggc agcgtggtcg aacggctaca caccgggatg tcgaggcctg cagcaaacga
     6181 cgaaaactac gctgcagcag tttagacaca tggcatggat gaactataca aataacccgg
     6241 gggatcccat ggtacgcgtg ctagaggcat caaataaaac gaaaggctca gtcgaaagac
     6301 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     6361 ccgccctaga cctag
//
