LOCUS       sequence-12663-e        7415 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 7415)
  AUTHORS   Dunlop MJ, Cox RS, Levine JH, Murray RM, Elowitz MB
  TITLE     Regulatory activity revealed by dynamic correlations in gene 
            expression noise.
  JOURNAL   Nat Genet. 2008 Dec . 40(12):1493-8.
  PUBMED    19029898
REFERENCE   2  (bases 1 to 7415)
  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: "Nat Genet. 
            2008 Dec . 40(12):1493-8."
COMMENT     Sequence Label: pNS2-sigmaVL
FEATURES             Location/Qualifiers
     source          1..7415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1..34)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     RBS             102..113
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    complement(854..887)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      896..967
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      983..1010
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1999..2010
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      complement(2351..2382)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     protein_bind    2396..2412
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    2419..2435
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             2483..2494
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2501..3211
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     CDS             3218..3931
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      3961..4047
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4582..5532)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      5580..5802
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(6294..6388)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6419..7213)
                     /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(6530..6549)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7140..7159
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7392..7410
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ataacttcgt atagcataca ttatacgaag ttatctcgag aataattctt gacatttatg
       61 cttccggctc gtataatacg tgcaattgga tccttaactt taaagaggag aaaggtaccg
      121 catgactagc aaaagaagca aaggtgaaga actgttcact ggtgttgttc caattctggt
      181 tgaactggat ggtgatgtta atggtcacaa attttctgtc tctggtgagg gtgaaggtga
      241 tgcaacctac ggtaaactga ccctgaaatt tatttgcact actggtaaac tgcctgttcc
      301 gtggccaacc ctggtcacta ctctgacttg gggtgttcaa tgctttgctc gttacccaga
      361 tcacatgaaa cagcatgact ttttcaagtc tgccatgccg gaaggttatg ttcaggaacg
      421 tactatcttt ttcaaagatg acggtaacta caagacccgt gctgaagtca agtttgaagg
      481 tgataccctg gttaatcgta tcgagctgaa aggtattgat tttaaagaag atggtaacat
      541 tctgggtcac aaactggaat acaacgctat ttctgataat gtatacatca ctgctgacaa
      601 acaaaagaat ggtatcaaag ctaatttcaa aattcgtcac aacattgaag atggtagcgt
      661 tcaactggca gaccattatc aacaaaatac tccaattggc gatggccctg tcctgctgcc
      721 agacaaccat tacctgtcca cccaatctcg tctgtctaaa gatccgaacg aaaagcgcga
      781 tcacatggtc ctgctggagt ttgtaaccgc tgctggtatt accctgggca tggatgaact
      841 gtataaataa tagataactt cgtatagcat acattatacg aagttatcca ggcatcaaat
      901 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
      961 cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttaacagct
     1021 attgtagata agtggggata ctacgggtat aaaaaaaccc gccggggcgg gtttttttac
     1081 gttgcttcag agctgatgtg gtggagatcg aatttcaaag gttagttttt tcatgacttc
     1141 cctctccccc aaataaaaag gcctgctggc gattaccagc cagcaggcct agtattaacc
     1201 gtgtccttct cgatttgaat aaataaaaac agccgttgcc agaaagaggc acggctgttt
     1261 ttattttcta ttatttgtac agctcatcca tgccaccggt agaatgacga ccctccgcgc
     1321 gctcatattg ctctacgatc gtataatctt cattatgaga ggtgatgtcc agtttaatat
     1381 tcacattgta cgcgccaggc agctgcacag gtttcttggc tttgtacgtg gttttcactt
     1441 cagcgtcata atggccgcca tctttcagtt tcaggcgctg tttaatttcg cctttcagcg
     1501 caccatcttc cggatacatg cgttcgctag acgcctccca acccatcgtc tttttctgca
     1561 tcaccgggcc atcagatgga aaattagtac cacgcagttt aactttatag atgaactcgc
     1621 catcttgcag ggaggagtcc tgagtgacgg tcacgacacc accatcttca aaattcatta
     1681 cgcgttccca cttgaaacct tccggaaaag acagcttcag atagtccggg atatccgctg
     1741 ggtgtttaac atacgcttta gaaccgtaca taaattgcgg gctcaggatg tcccacgcaa
     1801 aaggcagcgg gccgccttta gtcactttca gtttggcggt ctgggtgcct tcatacggac
     1861 ggccctcgcc ttcgccttcg atctcgaact cgtgaccgtt aacagaaccc tccatgtgaa
     1921 ctttgaagcg catgaactct ttaatgatag ccatgttatc ctcctcgccc ttggatcttt
     1981 tgctagtcat gcggtacctt tctcctcttt aaagttaaag atctccggac atgcaaccat
     2041 tatcaccgcc agaggtaaaa tagtcaacaa gcacggtgtt agatatttat cccttgcggt
     2101 gatagattta acgtatggtc gacgatccgg caaacaaacc accgttggta gcggtggttt
     2161 ttttgtttgg atcgacaatc ttcgtaagcg tcatcaataa gcgtaaaaaa accgggcaat
     2221 gcccggtttt ttaatgagaa attttacctg tcgtagccgc caccatccgg caaagaagca
     2281 tacaaggctt ttggcttata gctacgtagc gcattgcgtc gcagcacaat cccggcaccg
     2341 atcgagtaat agtcaaaagc ctccggtcgg aggcttttga ctttctgccc gggaattgtg
     2401 agcggataac aattgacatt gtgagcggat aacaagatac tgagcacatc agcaggacgc
     2461 actgaccgct agcgaattca ttaaagagga gaaaggtacc atgagcacaa aaaagaaacc
     2521 attaacacaa gagcagcttg aggacgcacg tcgccttaaa gcaatttatg aaaaaaagaa
     2581 aaatgaactt ggcttatccc aggaatctgt cgcagacaag atggggatgg ggcagtcagg
     2641 cgttggtgct ttatttaatg gcatcaatgc attaaatgct tataacgccg cattgcttgc
     2701 aaaaattctc aaagttagcg ttgaagaatt tagcccttca atcgccagag aaatctacga
     2761 gatgtatgaa gcggttagta tgcagccgtc acttagaagt gagtatgagt accctgtttt
     2821 ttctcatgtt caggcaggga tgttctcacc tgagcttaga acctttacca aaggtgatgc
     2881 ggagagatgg gtaagcacaa ccaaaaaagc cagtgattct gcattctggc ttgaggttga
     2941 aggtaattcc atgaccgcac caacaggctc caagccaagc tttcctgacg gaatgttaat
     3001 tctcgttgac cctgagcagg ctgttgagcc aggtgatttc tgcatagcca gacttggggg
     3061 tgatgagttt accttcaaga aactgatcag ggatagcggt caggtgtttt tacaaccact
     3121 aaacccacag tacccaatga tcccatgcaa tgagagttgt tccgttgtgg ggaaagttat
     3181 cgctagtcag tggcctgaag agacgtttgg gtaccgcatg cgtaaaggag aagaactttt
     3241 cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc
     3301 tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta aatttatttg
     3361 cactactgga aaactacctg ttccatggcc aacacttgtc actactttcg gttatggtct
     3421 aaaatgcttt gctagatacc cagatcatat gaaacggcat gactttttca agagtgccat
     3481 gcccgaaggt tatgtacagg aaagaactat atttttcaaa gatgacggga actacaagac
     3541 acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat agaatcgagt taaaaggtat
     3601 tgattttaaa gaagatggaa acattcttgg acacaaattg gaatacaact ataactcaca
     3661 caatgtatac atcatggcag acaaacaaaa gaatggaatc aaagttaact tcaaaattag
     3721 acacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa atactccaat
     3781 tggcgatggc cctgtccttt taccagacaa ccattacctg tcctatcaat ctgccctttc
     3841 gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa cagctgctgg
     3901 gattacacat ggcatggatg aactatacaa ataagcttaa ttagctgagt ctagaggcat
     3961 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     4021 gtgaacgctc tcctgagtag gacaaatccg ccgccctaga cctagggtac gggttttgct
     4081 gcccgcaaac gggctgttct ggtgttgcta gtttgttatc agaatcgcag atccggcttc
     4141 aggtttgccg gctgaaagcg ctatttcttc cagaattgcc atgatttttt ccccacggga
     4201 ggcgtcactg gctcccgtgt tgtcggcagc tttgattcga taagcagcat cgcctgtttc
     4261 aggctgtcta tgtgtgactg ttgagctgta acaagttgtc tcaggtgttc aatttcatgt
     4321 tctagttgct ttgttttact ggtttcacct gttctattag gtgttacatg ctgttcatct
     4381 gttacattgt cgatctgttc atggtgaaca gctttaaatg caccaaaaac tcgtaaaagc
     4441 tctgatgtat ctatcttttt tacaccgttt tcatctgtgc atatggacag ttttcccttt
     4501 gatatctaac ggtgaacagt tgttctactt ttgtttgtta gtcttgatgc ttcactgata
     4561 gatacaagag ccataagaac ctcagatcct tccgtattta gccagtatgt tctctagtgt
     4621 ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg agatcatgct tactttgcat
     4681 gtcactcaaa aattttgcct caaaactggt gagctgaatt tttgcagtta aagcatcgtg
     4741 tagtgttttt cttagtccgt tacgtaggta ggaatctgat gtaatggttg ttggtatttt
     4801 gtcaccattc atttttatct ggttgttctc aagttcggtt acgagatcca tttgtctatc
     4861 tagttcaact tggaaaatca acgtatcagt cgggcggcct cgcttatcaa ccaccaattt
     4921 catattgctg taagtgttta aatctttact tattggtttc aaaacccatt ggttaagcct
     4981 tttaaactca tggtagttat tttcaagcat taacatgaac ttaaattcat caaggctaat
     5041 ctctatattt gccttgtgag ttttcttttg tgttagttct tttaataacc actcataaat
     5101 cctcatagag tatttgtttt caaaagactt aacatgttcc agattatatt ttatgaattt
     5161 ttttaactgg aaaagataag gcaatatctc ttcactaaaa actaattcta atttttcgct
     5221 tgagaacttg gcatagtttg tccactggaa aatctcaaag cctttaacca aaggattcct
     5281 gatttccaca gttctcgtca tcagctctct ggttgcttta gctaatacac cataagcatt
     5341 ttccctactg atgttcatca tctgagcgta ttggttataa gtgaacgata ccgtccgttc
     5401 tttccttgta gggttttcaa tcgtggggtt gagtagtgcc acacagcata aaattagctt
     5461 ggtttcatgc tccgttaagt catagcgact aatcgctagt tcatttgctt tgaaaacaac
     5521 taattcagac atacatctca attggtctag gtgattttaa tcactatacc aattgagatg
     5581 ggctagtcaa tgataattac tagtcctttt cctttgagtt gtgggtatct gtaaattctg
     5641 ctagaccttt gctggaaaac ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc
     5701 tttgtgtgtt ttttttgttt atattcaagt ggttataatt tatagaataa agaaagaata
     5761 aaaaaagata aaaagaatag atcccagccc tgtgtataac tcactacttt agtcagttcc
     5821 gcagtattac aaaaggatgt cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa
     5881 ccctaaaggc ttaagtagca ccctcgcaag ctcgggcaaa tcgctgaata ttccttttgt
     5941 ctccgaccat caggcacctg agtcgctgtc tttttcgtga cattcagttc gctgcgctca
     6001 cggctctggc agtgaatggg ggtaaatggc actacaggcg ccttttatgg attcatgcaa
     6061 ggaaactacc cataatacaa gaaaagcccg tcacgggctt ctcagggcgt tttatggcgg
     6121 gtctgctatg tggtgctatc tgactttttg ctgttcagca gttcctgccc tctgattttc
     6181 cagtctgacc acttcggatt atcccgtgac aggtcattca gactggctaa tgcacccagt
     6241 aaggcagcgg tatcatcaac aggcttaccc gtcttactgt ccctagtgct tggattctca
     6301 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag
     6361 gtcattactg gatctatcaa caggagtcca agcgagctct cgaaccccag agtcccgctc
     6421 agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac
     6481 cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg
     6541 tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc
     6601 cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga
     6661 cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg gctggcgcga
     6721 gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac
     6781 gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg
     6841 tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag
     6901 atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag
     6961 tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg
     7021 ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg
     7081 ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg
     7141 cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat
     7201 cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagatcat gatcccctgc
     7261 gccatcagat ccttggcggc aagaaagcca tccagtttac tttgcagggc ttcccaacct
     7321 taccagaggg cgccccagct ggcaattccg acgtctaaga aaccattatt atcatgacat
     7381 taacctataa aaataggcgt atcacgaggc ccttt
//
