LOCUS       sequence-149519-        5086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcriptional interference assessment; strong interfering 
            promoter, landing site for spacers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5086)
  AUTHORS   Hoffmann SA, Kruse SM, Arndt KM
  TITLE     Long-range transcriptional interference in E. coli used to construct
            a dual positive selection system for genetic switches.
  JOURNAL   Nucleic Acids Res. 2016 Jun 2;44(10):e95. doi: 10.1093/nar/gkw125. 
            Epub 2016 Feb 29.
  PUBMED    26932362
REFERENCE   2  (bases 1 to 5086)
  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: "Nucleic 
            Acids Res."; date: "2016-06-2"; volume: "44(10):e95. doi"; pages: " 
            10.1093/nar/gkw125. Epub 2016 Feb 29"
COMMENT     Sequence Label: SAH0141-0.0-conP01
FEATURES             Location/Qualifiers
     source          1..5086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(202..1152)
                     /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      1200..1422
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(1914..2008)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(2164..2193)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     promoter        join(2248..2253,2254..2269,2270..2276)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             2311..3012
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             complement(3034..3744)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3312..3331)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3581..3599
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      3857..3888
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gttacattgt cgatctgttc atggtgaaca gctttaaatg caccaaaaac tcgtaaaagc
       61 tctgatgtat ctatcttttt tacaccgttt tcatctgtgc atatggacag ttttcccttt
      121 gatatctaac ggtgaacagt tgttctactt ttgtttgtta gtcttgatgc ttcactgata
      181 gatacaagag ccataagaac ctcagatcct tccgtattta gccagtatgt tctctagtgt
      241 ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg agatcatgct tactttgcat
      301 gtcactcaaa aattttgcct caaaactggt gagctgaatt tttgcagtta aagcatcgtg
      361 tagtgttttt cttagtccgt tacgtaggta ggaatctgat gtaatggttg ttggtatttt
      421 gtcaccattc atttttatct ggttgttctc aagttcggtt acgagatcca tttgtctatc
      481 tagttcaact tggaaaatca acgtatcagt cgggcggcct cgcttatcaa ccaccaattt
      541 catattgctg taagtgttta aatctttact tattggtttc aaaacccatt ggttaagcct
      601 tttaaactca tggtagttat tttcaagcat taacatgaac ttaaattcat caaggctaat
      661 ctctatattt gccttgtgag ttttcttttg tgttagttct tttaataacc actcataaat
      721 cctcatagag tatttgtttt caaaagactt aacatgttcc agattatatt ttatgaattt
      781 ttttaactgg aaaagataag gcaatatctc ttcactaaaa actaattcta atttttcgct
      841 tgagaacttg gcatagtttg tccactggaa aatctcaaag cctttaacca aaggattcct
      901 gatttccaca gttctcgtca tcagctctct ggttgcttta gctaatacac cataagcatt
      961 ttccctactg atgttcatca tctgagcgta ttggttataa gtgaacgata ccgtccgttc
     1021 tttccttgta gggttttcaa tcgtggggtt gagtagtgcc acacagcata aaattagctt
     1081 ggtttcatgc tccgttaagt catagcgact aatcgctagt tcatttgctt tgaaaacaac
     1141 taattcagac atacatctca attggtctag gtgattttaa tcactatacc aattgagatg
     1201 ggctagtcaa tgataattac atgtcctttt cctttgagtt gtgggtatct gtaaattctg
     1261 ctagaccttt gctggaaaac ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc
     1321 tttgtgtgtt ttttttgttt atattcaagt ggttataatt tatagaataa agaaagaata
     1381 aaaaaagata aaaagaatag atcccagccc tgtgtataac tcactacttt agtcagttcc
     1441 gcagtattac aaaaggatgt cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa
     1501 ccctaaaggc ttaagtagca ccctcgcaag ctcgggcaaa tcgctgaata ttccttttgt
     1561 ctccgaccat caggcacctg agtcgctgtc tttttcgtga cattcagttc gctgcgctca
     1621 cggctctggc agtgaatggg ggtaaatggc actacaggcg ccttttatgg attcatgcaa
     1681 ggaaactacc cataatacaa gaaaagcccg tcacgggctt ctcagggcgt tttatggcgg
     1741 gtctgctatg tggtgctatc tgactttttg ctgttcagca gttcctgccc tctgattttc
     1801 cagtctgacc acttcggatt atcccgtgac aggtcattca gactggctaa tgcacccagt
     1861 aaggcagcgg tatcatcaac aggcttaccc gtcttactgt ccctagtgct tggattctca
     1921 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag
     1981 gtcattactg gatctatcaa caggagtcca agcgagctcg taaacttggt ctgacagctc
     2041 tagctccggc aaaaaaacgg gcaaggtgtc accaccctgc cctttttctt taaaaccgaa
     2101 aagattactt cgcgtttgcc acctgacgtc taagaaaagg aatattcagc aatttgcccg
     2161 tgccgaagaa aggcccaccc gtgaaggtga gccagtgagt tgattgctac gtaattagtt
     2221 agttagccct tagtgactcg aattctcttg acaattaatc atcggctcgt ataatgtgtg
     2281 gaatctagag tcacacagga aacctactag atgagcgagc tgattaagga gaacatgcac
     2341 atgaagctgt acatggaggg caccgtggac aaccatcact tcaagtgcac atccgagggc
     2401 gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct
     2461 ctccccttcg ccttcgacat cctggctact agcttcctct acggcagcaa gaccttcatc
     2521 aaccacaccc agggcatccc cgacttcttc aagcagtcct tccctgaggg cttcacatgg
     2581 gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc
     2641 caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttcac atccaacggc
     2701 cctgtgatgc agaagaaaac actcggctgg gaggccttca ccgagacgct gtaccccgct
     2761 gacggcggcc tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg gagccatctg
     2821 atcgcaaaca tcaagaccac atatagatcc aagaaacccg ctaagaacct caagatgcct
     2881 ggcgtctact atgtggacta cagactggaa agaatcaagg aggccaacaa cgagacctac
     2941 gtcgagcagc acgaggtggc agtggccaga tactgcgacc tccctagcaa actggggcac
     3001 aagctaaatt aaaagctaac cggtcagaag cttttacttg tacagctcgt ccatgccgcc
     3061 ggtggagtgg cggccctcgg cgcgttcgta ctgttccacg atggtgtagt cctcgttgtg
     3121 ggaggtgatg tccaacttga tgttgacgtt gtaggcgccg ggcagctgca cgggcttctt
     3181 ggccttgtag gtggtcttga cctcagcgtc gtagtggccg ccgtccttca gcttcagcct
     3241 ctgcttgatc tcgcccttca gggcgccgtc ctcggggtac atccgctcgg aggaggcctc
     3301 ccagcccatg gtcttcttct gcattacggg gccgtcggag gggaagttgg tgccgcgcag
     3361 cttcaccttg tagatgaact cgccgtcctg caaggaggag tcctgggtca cggtcaccac
     3421 gccgccgtcc tcgaagttca tcacgcgctc ccacttgaag ccctcgggga aggacagctt
     3481 caagtagtcg gggatgtcgg cggggtgctt cacgtaggcc ttggagccgt acatgaactg
     3541 aggggacagg atgtcccagg cgaagggcag ggggccaccc ttggtcacct tcagcttggc
     3601 ggtctgggtg ccctcgtagg ggcggccctc gccctcgccc tcgatctcga actcgtggcc
     3661 gttcacggag ccctccatgt gcaccttgaa gcgcatgaac tccttgatga tggccatgtt
     3721 atcctcctcg cccttgctca ccatttatta cctcccgcct gattaatatt gaaactaacc
     3781 agggttattg tctcatgagc ggatacatat ttgaactgca ggagtcacta agggttagtt
     3841 agttagatta gcagaaagtc aaaagcctcc gaccggaggc ttttgactaa aacttccctt
     3901 ggggttatca ttggggctca ctcaaaggcg gtaatcagat aaaaaaaatc cttagctttc
     3961 gctaaggatg atttctgcta gtgtaagagg ttccaacttt caccataatg aaataagatc
     4021 actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgcg
     4081 cttacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt
     4141 tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca
     4201 gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca
     4261 gcagggcagt cgccctaaaa caaagttaaa catcatgagg gaagcggtga tcgccgaagt
     4321 atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct
     4381 ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga
     4441 tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga
     4501 ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac
     4561 cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt
     4621 tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat
     4681 tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc
     4741 ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac
     4801 cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac
     4861 gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc
     4921 cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca
     4981 ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt
     5041 tgtccactac gtgaaaggcg agatcaccaa ggtaataggt aaataa
//
