LOCUS       sequence-149523-        5586 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcriptional interference assessment; strong interfering 
            promoter, 500 bp spacer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5586)
  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 5586)
  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: SAH0146-0.5-conP01
FEATURES             Location/Qualifiers
     source          1..5586
                     /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"
     primer_bind     complement(3088..3107)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             complement(3534..4244)
                     /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(3812..3831)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4081..4099
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      4357..4388
                     /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 cggggagaaa aaaatcactg gatataccac cgttgatata
     3061 tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat
     3121 aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac
     3181 aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca tccggagttc
     3241 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
     3301 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
     3361 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
     3421 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc
     3481 accagttttg atttaaacgt ggccaatatg gacaacttac cggtcagaag cttttacttg
     3541 tacagctcgt ccatgccgcc ggtggagtgg cggccctcgg cgcgttcgta ctgttccacg
     3601 atggtgtagt cctcgttgtg ggaggtgatg tccaacttga tgttgacgtt gtaggcgccg
     3661 ggcagctgca cgggcttctt ggccttgtag gtggtcttga cctcagcgtc gtagtggccg
     3721 ccgtccttca gcttcagcct ctgcttgatc tcgcccttca gggcgccgtc ctcggggtac
     3781 atccgctcgg aggaggcctc ccagcccatg gtcttcttct gcattacggg gccgtcggag
     3841 gggaagttgg tgccgcgcag cttcaccttg tagatgaact cgccgtcctg caaggaggag
     3901 tcctgggtca cggtcaccac gccgccgtcc tcgaagttca tcacgcgctc ccacttgaag
     3961 ccctcgggga aggacagctt caagtagtcg gggatgtcgg cggggtgctt cacgtaggcc
     4021 ttggagccgt acatgaactg aggggacagg atgtcccagg cgaagggcag ggggccaccc
     4081 ttggtcacct tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc gccctcgccc
     4141 tcgatctcga actcgtggcc gttcacggag ccctccatgt gcaccttgaa gcgcatgaac
     4201 tccttgatga tggccatgtt atcctcctcg cccttgctca ccatttatta cctcccgcct
     4261 gattaatatt gaaactaacc agggttattg tctcatgagc ggatacatat ttgaactgca
     4321 ggagtcacta agggttagtt agttagatta gcagaaagtc aaaagcctcc gaccggaggc
     4381 ttttgactaa aacttccctt ggggttatca ttggggctca ctcaaaggcg gtaatcagat
     4441 aaaaaaaatc cttagctttc gctaaggatg atttctgcta gtgtaagagg ttccaacttt
     4501 caccataatg aaataagatc actaccgggc gtattttttg agttatcgag attttcagga
     4561 gctaaggaag ctaaaatgcg cttacgcaac tggtccagaa ccttgaccga acgcagcggt
     4621 ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct
     4681 atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg
     4741 agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa catcatgagg
     4801 gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc
     4861 catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg
     4921 aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg
     4981 cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt
     5041 ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca
     5101 gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc
     5161 gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc
     5221 gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta
     5281 tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat
     5341 gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc
     5401 gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc
     5461 gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc
     5521 gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtaataggt
     5581 aaataa
//
