LOCUS       62056_22540        4313 bp DNA     circular SYN 27-DEC-2020
DEFINITION  Cloning vector pVER-YFP-lacI, complete sequence.
ACCESSION   MT702634
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4313)
  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 4313)
  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 4313)
  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..4313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             101..913
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      1005..1549
                     /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."
     regulatory      complement(1996..2112)
                     /label=araC terminator
                     /note="araC terminator"
                     /regulatory_class="terminator"
     CDS             complement(2137..3216)
                     /label=lacI
                     /note="lac repressor"
     regulatory      complement(3245..3298)
                     /label=PlacI promoter
                     /note="PlacI promoter"
                     /regulatory_class="promoter"
     regulatory      complement(3303..3363)
                     /label=L3S2P21 terminator
                     /note="L3S2P21 terminator"
                     /regulatory_class="terminator"
     CDS             complement(3367..4083)
                     /label=EYFP
                     /note="enhanced YFP"
     misc_RNA        complement(4133..4183)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     protein_bind    complement(4187..4203)
                     /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)."
     promoter        complement(4211..4239)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     regulatory      complement(4261..4313)
                     /label=L3S3P21 terminator
                     /note="L3S3P21 terminator"
                     /regulatory_class="terminator"
ORIGIN
        1 ttgtgtctca aaatctctga tgtacattgc acaagataaa aatatatcat catgaacaat
       61 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
      121 aacgtcttgc tcgaggccgc 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 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc accactgatt
     1501 tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa acggctttgc
     1561 cgcggccctc tcacttccct gttaagtatc ttcctggcat cttccaggaa atctccgccc
     1621 cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga gtcagtgagc
     1681 gaggaagcgg aatatatcct gtatcacata ttctgctgac gcaccggtgc agcctttttt
     1741 ctcctgccac atgaagcact tcactgacac cctcatcagt gccaacatag taagccagta
     1801 tacactccgc tagcgctgag gtctgcctcg tgcagcgagt cagtgagcga ggaagcacct
     1861 cagataaaat atttgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     1921 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     1981 ccggcgtaga ggatctgctc atgtttgaca gcttatcatc gatgcataat gtgcctgtca
     2041 aatggacgaa gcagggattc tgcaaaccct atgctactcc ctcgagccgt caattgtctg
     2101 attcgttacc aattattttt tcctcctgga ttatcactgc ccgctttcca gtcgggaaac
     2161 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtaat
     2221 gggcgccagg gtggtttttc ttttcaccag tgagactggc aacagctgat tgcccttcac
     2281 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     2341 atcctgtttg atggtggtta acggcgggat ataacatgag ctatcttcgg tatcgtcatg
     2401 tcccactacc gagatattcg caccaacgcg cagcccggac tcgataatgg cgcgcattgc
     2461 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     2521 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     2581 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     2641 ggcagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     2701 ctccacgccc agtcgcgtac cgtcctcatg ggagaaaata atactgttga tgggtgtctg
     2761 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     2821 atcctgctca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     2881 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     2941 ggcacccagt tgatcggcgc gagatttaat cgccgcgacg atttgcgacg gcgcgtgcag
     3001 ggccaggctg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca ggtgttgtgc
     3061 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     3121 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     3181 atactctgcg acatcgtata acgttactgg tttcatattc accaccctga attgactctc
     3241 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg cgcgccgcca
     3301 ttggaccaaa acgaaaaaag gccccccttt cgggaggcct cttttctgga atttggtacc
     3361 gagttacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag
     3421 caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact ggtagctaag
     3481 gtagtggttg tcgggcagca gcacggggcc gtcgccgatt ggggtgttct gctggtagtg
     3541 gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat
     3601 gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag
     3661 cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt
     3721 caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt
     3781 gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg
     3841 cagcttcatg tggtcggggt agcgggcgaa gcattgcagg ccgtagccga aggtggtcac
     3901 gagggtgggc cagggcacgg gcagcttgcc tgtggtgcag atgaacttca gggtcagctt
     3961 gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc
     4021 gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac
     4081 catctagtat ttcccctctt tctctagtat taaacaaaat tatttgtaga ggctgtttcg
     4141 tcctcacgga ctcatcagac cggaaagcac atccggtgac agctaattgt tatccgctca
     4201 caattccaca cattatacga gccgatgatt aattgtcaac acagccaacg atcgttaagc
     4261 gggagaccag aaacaaaaaa aggccccccg ttagggaggc cttcaataat tgg
//
