LOCUS       V016035                 5693 bp    DNA     circular SYN 28-JUL-2021
DEFINITION  Exported.
ACCESSION   V016035
VERSION     V016035
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5693)
  AUTHORS   Becker K, Meyer A, Roberts TM, Panke S.
  TITLE     Plasmid replication based on the T7 origin of replication requires a
            T7 RNAP variant and inactivation of ribonuclease H
  JOURNAL   Nucleic Acids Res (2021) In press
   PUBMED   34255845
REFERENCE   2  (bases 1 to 5693)
  AUTHORS   Becker K.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-JUN-2021) D-BSSE, ETHZ, Mattenstrasse 26, Basel, BS
            4058, Switzerland
REFERENCE   3  (bases 1 to 5693)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
            Acids Res (2021) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (10-JUN-2021) D-BSSE, ETHZ, Mattenstrasse 26, Basel, BS 4058,
            Switzerland"
FEATURES             Location/Qualifiers
     source          1..5693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        192..220
                     /label="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    228..244
                     /label="lac operator"
                     /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)."
     CDS             287..2935
                     /note="T7 RNA polymerase from Escherichia phage T7.
                     Accession#: P00573"
                     /label="T7 RNA polymerase"
     CDS             complement(2958..3749)
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     CDS             complement(4237..5184)
                     /label="Rep101"
                     /note="RepA protein needed for replication with the pSC101
                     origin"
     rep_origin      complement(5232..5454)
                     /direction=LEFT
                     /label="pSC101 ori"
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 aggatccgga gcttatcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg
       61 gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg
      121 cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct
      181 gaaatgagct gttgacaatt aatcatcggc tcgtataatg tgtggaattg tgagcggata
      241 acaatttcac acaggaaaca gaattcctaa ctggaagagg cactaaatga acacgattaa
      301 catcgctaag aacgacttct ctgacatcga actggctgct atcccgttca acactctggc
      361 tgaccattac ggtgagcgtt tagctcgcga acagttggcc cttgagcatg agtcttacga
      421 gatgggtgaa gcacgcttcc gcaagatgtt tgagcgtcaa cttaaagctg gtgaggttgc
      481 ggataacgct gccgccaagc ctctcatcac taccctactc cctaagatga ttgcacgcat
      541 caacgactgg tttgaggaag tgaaagctaa gcgcggcaag cgcccgacag ccttccagtt
      601 cctgcaagaa atcaagccgg aagccgtagc gtacatcacc attaagacca ctctggcttg
      661 cctaaccagt gctgacaata caaccgttca ggctgtagca agcgcaatcg gtcgggccat
      721 tgaggacgag gctcgcttcg gtcgtatccg tgaccttgaa gctaagcact tcaagaaaaa
      781 cgttgaggaa caactcaaca agcgcgtagg gcacgtctac aagaaagcat ttatgcaagt
      841 tgtcgaggct gacatgctct ctaagggtct actcggtggc gaggcgtggt cttcgtggca
      901 taaggaagac tctattcatg taggagtacg ctgcatcgag atgctcattg agtcaaccgg
      961 aatggttagc ttacaccgcc aaaatgctgg cgtagtaggt caagactctg agactatcga
     1021 actcgcacct gaatacgctg aggctatcgc aacccgtgca ggtgcgctgg ctggcatctc
     1081 tccgatgttc caaccttgcg tagttcctcc taagccgtgg actggcatta ctggtggtgg
     1141 ctattgggct aacggtcgtc gtcctctggc gctggtgcgt actcacagta agaaagcact
     1201 gatgcgctac gaagacgttt acatgcctga ggtgtacaaa gcgattaaca ttgcgcaaaa
     1261 caccgcatgg aaaatcaaca agaaagtcct agcggtcgcc aacgtaatca ccaagtggaa
     1321 gcattgtccg gtcgaggaca tccctgcgat tgagcgtgaa gaactcccga tgaaaccgga
     1381 agacatcgac atgaatcctg aggctctcac cgcgtggaaa cgtgctgccg ctgctgtgta
     1441 ccgcaaggac aaggctcgca agtctcgccg tatcagcctt gagttcatgc ttgagcaagc
     1501 caataagttt gctaaccata aggccatctg gttcccttac aacatggact ggcgcggtcg
     1561 tgtttacgct gtgtcaatgt tcaacccgca aggtaacgat atgaccaaag gactgcttac
     1621 gctggcgaaa ggtaaaccaa tcggtaagga aggttactac tggctgaaaa tccacggtgc
     1681 aaactgtgcg ggtgtcgata aggttccgtt ccctgagcgc atcaagttca ttgaggaaaa
     1741 ccacgagaac atcatggctt gcgctaagtc tccactggag aacacttggt gggctgagca
     1801 agattctccg ttctgcttcc ttgcgttctg ctttgagtac gctggggtac agcaccacgg
     1861 cctgagctat aactgctccc ttccgctggc gtttgacggg tcttgctctg gcatccagca
     1921 cttctccgcg atgctccgag atgaggtagg tggtcgcgcg gttaacttgc ttcctagtga
     1981 aaccgttcag gacatctacg ggattgttgc taagaaagtc aacgagattc tacaagcaga
     2041 cgcaatcaat gggaccgata acgaagtagt taccgtgacc gatgagaaca ctggtgaaat
     2101 ctctgagaaa gtcaagctgg gcactaaggc actggctggt caatggctgg cttacggtgt
     2161 tactcgcagt gtgactaagc gttcagtcat gacgctggct tacgggtcca aagagttcgg
     2221 cttccgtcaa caagtgctgg aagataccat tcagccagct attgattccg gcaagggtct
     2281 gatgttcact cagccgaatc aggctgctgg atacatggct aagctgattt gggaatctgt
     2341 gagcgtgacg gtggtagctg cggttgaagc aatgaactgg cttaagtctg ctgctaagct
     2401 gctggctgct gaggtcaaag ataagaagac tggagagatt cttcgcaagc gttgcgctgt
     2461 gcattgggta actcctgatg gtttccctgt gtggccggaa tacaagaagc ctattcagac
     2521 gcgcttgaac ctgatgttcc tcggtcagtt ccgcttacag cctaccatta acaccaacaa
     2581 agatagcgag attgatgcac acaaacagga gtctggtatc gctcctaact ttgtacacag
     2641 ccaagacggt agccaccttc gtaagactgt agtgtgggca cacgagaagt acggaatcga
     2701 atcttttgca ctgattcacg actccttcgg taccattccg gctgacgctg cgaacctgtt
     2761 caaagcagtg cgcgaaacta tggttgacac atatgagtct tgtgatgtac tggctgattt
     2821 ctacgaccag ttcgctgacc agttgcacga gtctcaattg gacaaaatgc cagcacttcc
     2881 ggctaaaggt aacttgaacc tccgtgacat cttagagtcg gacttcgcgt tcgcgtaacg
     2941 ccaagatccc cttattagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa
     3001 tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct
     3061 tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg
     3121 ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca
     3181 tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag cctggcgaac
     3241 agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg
     3301 gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag
     3361 gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg
     3421 gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag
     3481 tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc
     3541 agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga caggtcggtc
     3601 ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag
     3661 ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa
     3721 cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatca
     3781 gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca gtttactttg
     3841 cagggcttcc caaccttacc agagggcgcc ccagctggca attccggttc gcttgctgtc
     3901 cataaaaccg cccagtctag ctatcgccat gtaagcccac tgcaagctac ctgctttctc
     3961 tttgcgcttg cgttttccct tgtccagata gccctttcac ctgttctatt aggtgttaca
     4021 tgctgttcat ctgttacatt gtcgatctgt tcatggtgaa cagctttaaa tgcaccaaaa
     4081 actcgtaaaa gctctgatgt atctatcttt tttacaccgt tttcatctgt gcatatggac
     4141 agttttccct ttgatatgta acggtgaaca gttgttctac ttttgtttgt tagtcttgat
     4201 gcttcactga tagatacaag agccataaga acctcagatc cttccgtatt tagccagtat
     4261 gttctctagt gtggttcgtt gtttttgcgt gagccatgag aacgaaccat tgagatcata
     4321 cttactttgc atgtcactca aaaattttgc ctcaaaactg gtgagctgaa tttttgcagt
     4381 taaagcatcg tgtagtgttt ttcttagtcc gttatgtagg taggaatctg atgtaatggt
     4441 tgttggtatt ttgtcaccat tcatttttat ctggttgttc tcaagttcgg ttacgagatc
     4501 catttgtcta tctagttcaa cttggaaaat caacgtatca gtcgggcggc ctcgcttatc
     4561 aaccaccaat ttcatattgc tgtaagtgtt taaatcttta cttattggtt tcaaaaccca
     4621 ttggttaagc cttttaaact catggtagtt attttcaagc attaacatga acttaaattc
     4681 atcaaggcta atctctatat ttgccttgtg agttttcttt tgtgttagtt cttttaataa
     4741 ccactcataa atcctcatag agtatttgtt ttcaaaagac ttaacatgtt ccagattata
     4801 ttttatgaat ttttttaact ggaaaagata aggcaatatc tcttcactaa aaactaattc
     4861 taatttttcg cttgagaact tggcatagtt tgtccactgg aaaatctcaa agcctttaac
     4921 caaaggattc ctgatttcca cagttctcgt catcagctct ctggttgctt tagctaatac
     4981 accataagca ttttccctac tgatgttcat catctgagcg tattggttat aagtgaacga
     5041 taccgtccgt tctttccttg tagggttttc aatcgtgggg ttgagtagtg ccacacagca
     5101 taaaattagc ttggtttcat gctccgttaa gtcatagcga ctaatcgcta gttcatttgc
     5161 tttgaaaaca actaattcag acatacatct caattggtct aggtgatttt aatcactata
     5221 ccaattgaga tgggctagtc aatgataatt actagtcctt ttcctttgag ttgtgggtat
     5281 ctgtaaattc tgctagacct ttgctggaaa acttgtaaat tctgctagac cctctgtaaa
     5341 ttccgctaga cctttgtgtg ttttttttgt ttatattcaa gtggttataa tttatagaat
     5401 aaagaaagaa taaaaaaaga taaaaagaat agatcccagc cctgtgtata actcactact
     5461 ttagtcagtt ccgcagtatt acaaaaggat gtcgcaaacg ctgtttgctc ctctacaaaa
     5521 cagaccttaa aaccctaaag gcttaagtag caccctcgca agctcgggca aatcgctgaa
     5581 tattcctttt gtctccgacc atcaggcacc tgagtcgctg tctttttcgt gacattcagt
     5641 tcgctgcgct cacggctctg gcagtgaatg ggggtaaatg gcactacagg cgc
//
