LOCUS       sequence-266155-        5048 bp    DNA     circular SYN 15-APR-2025
DEFINITION  pZS2-pLsoxS-TEVp-proD-soxR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5048)
  AUTHORS   Muller IE, Rubens JR, Jun T, Graham D, Xavier R, Lu TK
  TITLE     Gene networks that compensate for crosstalk with crosstalk.
  JOURNAL   Nat Commun. 2019 Sep 6;10(1):4028. doi: 10.1038/s41467-019-12021-y.
  PUBMED    31492904
REFERENCE   2  (bases 1 to 5048)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2019-09-6"; volume: "10(1):4028. doi"; pages: " 
            10.1038/s41467-019-12021-y"
COMMENT     Sequence Label: pIM04
FEATURES             Location/Qualifiers
     source          1..5048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             80..91
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             101..808
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="GESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNK
                     HLFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQ
                     REERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFI
                     VGIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPE
                     EPFQPVKEATQLMN"
     terminator      862..948
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1639..1725
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2270..3220)
                     /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      3268..3490
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3988..4082)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4113..4907)
                     /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(4224..4243)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4834..4853
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 cccataaatc gctttacctc aagttaactt gaggaattat actccccaac atcagcagga
       61 cgcactgacc gaattcatta aagaggagaa aggtaccatg ggagaaagct tgtttaaggg
      121 gccgcgtgat tacaacccga tatcgagcac catttgtcat ttgacgaatg aatctgatgg
      181 gcacacaaca tcgttgtatg gtattggatt tggtcccttc atcattacaa acaagcactt
      241 gtttagaaga aataatggaa cactgttggt ccaatcacta catggtgtat tcaaggtcaa
      301 gaacaccacg actttgcaac aacacctcat tgatgggagg gacatgataa ttattcgcat
      361 gcctaaggat ttcccaccat ttcctcaaaa gctgaaattt agagagccac aaagggaaga
      421 gcgcatatgt cttgtgacaa ccaacttcca aactaagagc atgtctagca tggtgtcaga
      481 cactagttgc acattccctt catctgatgg catattctgg aagcattgga ttcaaaccaa
      541 ggatgggcag tgtggcagtc cattagtatc aactagagat gggttcattg ttggtataca
      601 ctcagcatcg aatttcacca acacaaacaa ttatttcaca agcgtgccga aaaacttcat
      661 ggaattgttg acaaatcagg aggcgcagca gtgggttagt ggttggcgat taaatgctga
      721 ctcagtattg tgggggggcc ataaagtttt catggtgaaa cctgaagagc cttttcagcc
      781 agttaaggaa gcgactcaac tcatgaatcg tcgtcgccgt cgctaacccg ggggatccca
      841 tggtacgcgt gctagaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
      901 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgccctag
      961 agctagccac agctaacacc acgtcgtccc tatctgctgc cctaggtcta tgagtggttg
     1021 ctggataact ttacgggcat gcataaggct cgtataatat attcagggag accacaacgg
     1081 tttccctcta caaataattt tgtttaactt tgaattcttc acacaggaaa ccggtaccat
     1141 ggaaaagaaa ttaccccgca ttaaagcgct gctaaccccc ggcgaagtgg cgaaacgcag
     1201 cggtgtggcg gtatcggcgc tgcatttcta tgaaagtaaa gggttgatta ccagtatccg
     1261 taacagcggc aatcagcggc gatataaacg tgatgtgttg cgatatgttg caattatcaa
     1321 aattgctcag cgtattggca ttccgctggc gaccattggt gaagcgtttg gcgtgttgcc
     1381 cgaagggcat acgttaagtg cgaaagagtg gaaacagctt tcgtcccaat ggcgagaaga
     1441 gttggatcgg cgcattcata ccttagtggc gctgcgtgac gaactggacg gatgtattgg
     1501 ttgtggctgc ctttcgcgca gtgattgccc gttgcgtaac ccgggcgacc gcttaggaga
     1561 agaaggtacc ggcgcacgct tgctggaaga tgaacaaaac taacccgggg gatcccatgg
     1621 tacgcgtgct agaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     1681 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gccctagacc
     1741 tagggcctag ggtacgggtt ttgctgcccg caaacgggct gttctggtgt tgctagtttg
     1801 ttatcagaat cgcagatccg gcttcagccg gtttgccggc tgaaagcgct atttcttcca
     1861 gaattgccat gattttttcc ccacgggagg cgtcactggc tcccgtgttg tcggcagctt
     1921 tgattcgata agcagcatcg cctgtttcag gctgtctatg tgtgactgtt gagctgtaac
     1981 aagttgtctc aggtgttcaa tttcatgttc tagttgcttt gttttactgg tttcacctgt
     2041 tctattaggt gttacatgct gttcatctgt tacattgtcg atctgttcat ggtgaacagc
     2101 tttgaatgca ccaaaaactc gtaaaagctc tgatgtatct atctttttta caccgttttc
     2161 atctgtgcat atggacagtt ttccctttga tatgtaacgg tgaacagttg ttctactttt
     2221 gtttgttagt cttgatgctt cactgataga tacaagagcc ataagaacct cagatccttc
     2281 cgtatttagc cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg
     2341 aaccattgag atcatactta ctttgcatgt cactcaaaaa ttttgcctca aaactggtga
     2401 gctgaatttt tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta tgtaggtagg
     2461 aatctgatgt aatggttgtt ggtattttgt caccattcat ttttatctgg ttgttctcaa
     2521 gttcggttac gagatccatt tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg
     2581 ggcggcctcg cttatcaacc accaatttca tattgctgta agtgtttaaa tctttactta
     2641 ttggtttcaa aacccattgg ttaagccttt taaactcatg gtagttattt tcaagcatta
     2701 acatgaactt aaattcatca aggctaatct ctatatttgc cttgtgagtt ttcttttgtg
     2761 ttagttcttt taataaccac tcataaatcc tcatagagta tttgttttca aaagacttaa
     2821 catgttccag attatatttt atgaattttt ttaactggaa aagataaggc aatatctctt
     2881 cactaaaaac taattctaat ttttcgcttg agaacttggc atagtttgtc cactggaaaa
     2941 tctcaaagcc tttaaccaaa ggattcctga tttccacagt tctcgtcatc agctctctgg
     3001 ttgctttagc taatacacca taagcatttt ccctactgat gttcatcatc tgagcgtatt
     3061 ggttataagt gaacgatacc gtccgttctt tccttgtagg gttttcaatc gtggggttga
     3121 gtagtgccac acagcataaa attagcttgg tttcatgctc cgttaagtca tagcgactaa
     3181 tcgctagttc atttgctttg aaaacaacta attcagacat acatctcaat tggtctaggt
     3241 gattttaatc actataccaa ttgagatggg ctagtcaatg ataattacta gtccttttcc
     3301 tttgagttgt gggtatctgt aaattctgct agacctttgc tggaaaactt gtaaattctg
     3361 ctagaccctc tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat attcaagtgg
     3421 ttataattta tagaataaag aaagaataaa aaaagataaa aagaatagat cccagccctg
     3481 tgtataactc actactttag tcagttccgc agtattacaa aaggatgtcg caaacgctgt
     3541 ttgctcctct acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc ctcgcaagct
     3601 cgggcaaatc gctgaatatt ccttttgtct ccgaccatca ggcacctgag tcgctgtctt
     3661 tttcgtgaca ttcagttcgc tgcgctcacg gctctggcag tgaatggggg taaatggcac
     3721 tacaggcgcc ttttatggat tcatgcaagg aaactaccca taatacaaga aaagcccgtc
     3781 acgggcttct cagggcgttt tatggcgggt ctgctatgtg gtgctatctg actttttgct
     3841 gttcagcagt tcctgccctc tgattttcca gtctgaccac ttcggattat cccgtgacag
     3901 gtcattcaga ctggctaatg cacccagtaa ggcagcggta tcatcaacag gcttacccgt
     3961 cttactgtcc ctagtactag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc
     4021 gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag
     4081 tccaagcgag ctctcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag
     4141 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
     4201 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg
     4261 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
     4321 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgcgc
     4381 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
     4441 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
     4501 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
     4561 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
     4621 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     4681 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg
     4741 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     4801 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     4861 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct
     4921 catcctgtct cttgatcaga tcttgatccc ctgcgccatc agatccttgg cggcaagaaa
     4981 gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat
     5041 tccgacgt
//
