LOCUS       62056_21685        5661 bp DNA     circular SYN 27-DEC-2020
DEFINITION  Cloning vector pTY1-lacI, complete sequence.
ACCESSION   MT702633
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5661)
  AUTHORS   Tack DS, Tonner PD, Pressman A, Olson ND, Levy SF, Romantseva EF, 
            Alperovich N, Vasilyeva O, Ross D.
  TITLE     The genotype-phenotype landscape of an allosteric protein
  JOURNAL   bioRxiv (2020) In press
REFERENCE   2  (bases 1 to 5661)
  AUTHORS   Tack DS.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-JUL-2020) Cellular Engineering, National Institute of 
            Standards and Technology, 100 Bureau Dr MS8313, Gaithersburg, MD 
            20899, USA
REFERENCE   3  (bases 1 to 5661)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            (2020) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (01-JUL-2020) Cellular Engineering, National Institute of Standards 
            and Technology, 100 Bureau Dr MS8313, Gaithersburg, MD 20899, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             101..913
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      1005..1550
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     terminator      complement(1904..1951)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      complement(2126..2242)
                     /label=araC terminator
                     /note="araC terminator"
                     /regulatory_class="terminator"
     CDS             complement(2267..3346)
                     /label=lacI
                     /note="lac repressor"
     regulatory      complement(3375..3428)
                     /label=Plac promoter
                     /note="Plac promoter"
                     /regulatory_class="promoter"
     promoter        3503..3531
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    3539..3555
                     /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)."
     misc_RNA        3559..3609
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             3659..4861
                     /label=TcR
                     /note="tetracycline efflux protein"
     CDS             4885..5601
                     /label=EYFP
                     /note="enhanced YFP"
     regulatory      complement(5609..5661)
                     /label=L3S3P21
                     /note="L3S3P21"
                     /regulatory_class="terminator"
ORIGIN
        1 ttgtgtctca aaatctctga tgtacattgc acaagataaa aatatatcat catgaacaat
       61 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
      121 aacgtcttgc tccaggccgc gattaaattc caacatggat gctgatttat atgggtataa
      181 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc
      241 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
      301 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
      361 tatccgtact cctgatgatg catggttact caccactgcg atccccggga aaacagcatt
      421 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
      481 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
      541 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
      601 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata agcttttgcc
      661 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
      721 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
      781 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
      841 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
      901 cgatgagttt ttctaaggta actgtcagac caagtttact catatatact ttagattgat
      961 ttttcgttcc actgagcgtc agacccctta ataagatgat cttcttgaga tcgttttggt
     1021 ctgcgcgtaa tctcttgctc tgaaaacgaa aaaaccgcct tgcagggcgg tttttcgaag
     1081 gttctctgag ctaccaactc tttgaaccga ggtaactggc ttggaggagc gcagtcacca
     1141 aaacttgtcc tttcagttta gccttaaccg gcgcatgact tcaagactaa ctcctctaaa
     1201 tcaattacca gtggctgctg ccagtggtgc ttttgcatgt ctttccgggt tggactcaag
     1261 acgatagtta ccggataagg cgcagcggtc ggactgaacg gggggttcgt gcatacagtc
     1321 cagcttggag cgaactgcct acccggaact gagtgtcagg cgtggaatga gacaaacgcg
     1381 gccataacag cggaatgaca ccggtaaacc gaaaggcagg aacaggagag cgcacgaggg
     1441 agccgccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc caccactgat
     1501 ttgagcgtca gatttcgtga tgcttgtcag gggggcggag cctatggaaa aacggctttg
     1561 ccgcggccct ctcacttccc tgttaagtat cttcctggca tcttccagga aatctccgcc
     1621 ccgttcgtaa gccatttccg ctcgccgcag tcgaacgacc gagcgtagcg agtcagtgag
     1681 cgaggaagcg gaatatatcc tgtatcacat attctgctga cgcaccggtg cagccttttt
     1741 tctcctgcca catgaagcac ttcactgaca ccctcatcag tgccaacata gtaagccagt
     1801 atacactccg ctagcgctga ggtctgcctc gtgcagcgag tcagtgagcg aggaagcacc
     1861 tcagataaaa tatttgctca tgagcccgaa gtggcgagcc cgacaaaaaa cccctcaaga
     1921 cccgtttaga ggccccaagg ggttatgcta gtcttcccca tcggtgagcc cgggctgtcg
     1981 gcgtnntnnn anntnnnann tnnnanntnn nanntnnnan natatgccag caggccggcc
     2041 acgctnntnn nanntnnnan nannnanntn nnanntnnna nncggtggcc cgggcggccg
     2101 cacgatgcgt ccggcgtaga ggatctgctc atgtttgaca gcttatcatc gatgcataat
     2161 gtgcctgtca aatggacgaa gcagggattc tgcaaaccct atgctactcc ctcgagccgt
     2221 caattgtctg attcgttacc aattattttt tcctcctgga ttatcactgc ccgctttcca
     2281 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2341 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagactggc aacagctgat
     2401 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
     2461 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctatcttcgg
     2521 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg
     2581 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
     2641 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
     2701 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
     2761 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
     2821 cgaccagatg ctccacgccc agtcgcgtac cgtcctcatg ggagaaaata atactgttga
     2881 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
     2941 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
     3001 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
     3061 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
     3121 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
     3181 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
     3241 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
     3301 agacaccggc atactctgcg acatcgtata acgttactgg tttcatattc accaccctga
     3361 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
     3421 cgcgccgcca taaatctcgg taccaaattc cagaaaagag gcctcccgaa aggggggcct
     3481 tttttcgttt tggtccgagc tgttgacaat taatcatcgg ctcgtataat gtgtggaatt
     3541 gtgagcggat aacaattagc tgtcaccgga tgtgctttcc ggtctgatga gtccgtgagg
     3601 acgaaacagc ctctacaaat aattttgttt aatactagcc atcaaggaga gctgctacat
     3661 gagtagcagt acgaaaattg cgcttgtcat caccctcctg gatgcgatgg ggatcggctt
     3721 gatcatgccg gtactgccaa cccttctgcg cgagttcatt gcaagcgaag atattgccaa
     3781 ccatttcggg gttctgctcg cactgtacgc cttaatgcag gtcatctttg ctccctggtt
     3841 aggcaaaatg tcagacagct ttggacgccg tcctgttttg ctgttaagcc ttatcggagc
     3901 gagcctggat taccttttat tggccttctc ctcggcactg tggatgcttt atttgggtcg
     3961 tttgctgagt gggattacag gcgcgacggg tgccgtggcg gcgtcggtga ttgctgatac
     4021 gacgtccgca agtcaacgtg tgaaattgtt cggctggtta ggagcctcct ttggcttggg
     4081 cttaatcgct gggccaatta ttggcgggtt cgccggcgaa atctcaccac attccccttt
     4141 tttcatcgcg gcattactca acattgtcac gttcctggtg gtgatgttct ggttccgcga
     4201 aacgaaaaac acccgcgata acacggatac agaggtgggg gttgaaacgc aatcgaacag
     4261 tgtgtacatc acgctcttca agaccatgcc catcctgctc atcatctact tctccgcaca
     4321 gttgattggg caaatcccgg ccacagtgtg ggttttgttt acggaaaacc gtttcgggtg
     4381 gaactccatg atggtgggtt tctctctggc tggattggga cttctgcata gtgttttcca
     4441 ggctttcgtc gctggccgta ttgccacaaa gtggggagaa aaaaccgctg tattgcttgg
     4501 ttttatcgca gatagctctg cgtttgcctt cttggcattt attagcgaag gctggctcgt
     4561 gtttccggta ttgattctgt tggctggggg cggtatcgca ttacccgcgc tgcagggagt
     4621 tatgtctatt caaaccaaat cacaccaaca aggagcgctg caaggcttac ttgtgtccct
     4681 gaccaacgca accggagtca tcgggccact tctgttcgct gtaatttata accactcact
     4741 gccaatttgg gatggatgga tctggatcat cggtcttgcc ttctactgca tcatcatttt
     4801 gctgtcaatg acattcatgc tgacgcctca agcccaagga tctaaacaag aaacgagtgc
     4861 ctaacggcgt aaggaggtat ttttatggtg tcaaagggtg aggaactgtt tacggggatc
     4921 gtcccgattc ttgttgaact tgacggcgac gtaaatggtc acaagttttc cgtatcgggc
     4981 gaaggtgagg gcgatgcgac ttatgggaaa ttaacactga aattcatttg caccaccgga
     5041 aaactgcccg ttccttggcc tactctggta accacgttcg gatatggttt acagtgtttt
     5101 gctcgctacc cggaccatat gaaactgcac gatttcttca agtccgccat gccggagggc
     5161 tacgtgcagg aacgtacaat cttcttcaaa gacgatggta attacaagac ccgtgctgaa
     5221 gttaaatttg agggggatac tttagtcaat cgtattgaat tgaaggggat tgactttaag
     5281 gaagacggta atatccttgg ccacaagctt gaatacaact acaatagtca caatgtgtat
     5341 attatggctg ataaacagaa gaatggcatt aaggttaact ttaagatccg tcacaatatc
     5401 gaagacggat ctgtccagct tgctgaccat tatcagcaaa atacgccgat cggtgacggt
     5461 ccggttttgt tgccggacaa ccattacctg tcctatcagt cggcgttaag taaagatccg
     5521 aatgagaaac gcgaccatat ggttttgttg gagtttgtga cggctgctgg cattacgctt
     5581 gggatggacg agctgtataa ataaaagcgg gagaccagaa acaaaaaaag gccccccgtt
     5641 agggaggcct tcaataattg g
//
