LOCUS       sequence-149522-        5071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcriptional interference assessment; no interfering promoter 
            (scrambled).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5071)
  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 5071)
  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: SAH0098-scrPX
FEATURES             Location/Qualifiers
     source          1..5071
                     /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"
     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(3019..3729)
                     /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(3297..3316)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3566..3584
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     terminator      3842..3873
                     /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 aattcacgta ccgatgtgtt taacaaattg gtcaggattt
     2281 atctctagag 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 aaaagctttt acttgtacag ctcgtccatg ccgccggtgg agtggcggcc
     3061 ctcggcgcgt tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg tgatgtccaa
     3121 cttgatgttg acgttgtagg cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt
     3181 cttgacctca gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct tgatctcgcc
     3241 cttcagggcg ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc ccatggtctt
     3301 cttctgcatt acggggccgt cggaggggaa gttggtgccg cgcagcttca ccttgtagat
     3361 gaactcgccg tcctgcaagg aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa
     3421 gttcatcacg cgctcccact tgaagccctc ggggaaggac agcttcaagt agtcggggat
     3481 gtcggcgggg tgcttcacgt aggccttgga gccgtacatg aactgagggg acaggatgtc
     3541 ccaggcgaag ggcagggggc cacccttggt caccttcagc ttggcggtct gggtgccctc
     3601 gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg tggccgttca cggagccctc
     3661 catgtgcacc ttgaagcgca tgaactcctt gatgatggcc atgttatcct cctcgccctt
     3721 gctcaccatt tattacctcc cgcctgatta atattgaaac taaccagggt tattgtctca
     3781 tgagcggata catatttgaa ctgcaggagt cactaagggt tagttagtta gattagcaga
     3841 aagtcaaaag cctccgaccg gaggcttttg actaaaactt cccttggggt tatcattggg
     3901 gctcactcaa aggcggtaat cagataaaaa aaatccttag ctttcgctaa ggatgatttc
     3961 tgctagtgta agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt
     4021 ttttgagtta tcgagatttt caggagctaa ggaagctaaa atgcgcttac gcaactggtc
     4081 cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta
     4141 tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc
     4201 cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg gcagtcgccc
     4261 taaaacaaag ttaaacatca tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc
     4321 agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta
     4381 cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc tggttacggt
     4441 gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc
     4501 ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga
     4561 cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg
     4621 caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt
     4681 gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt
     4741 tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa
     4801 ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg
     4861 gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga
     4921 gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca
     4981 agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa
     5041 aggcgagatc accaaggtaa taggtaaata a
//
