LOCUS       62056_13245        5295 bp DNA     circular SYN 22-JUL-2020
DEFINITION  Cloning vector pJV298-Tyr1, complete sequence.
ACCESSION   MT346028
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5295)
  AUTHORS   Lee MS, Hung CS, Phillips DA, Buck CC, Gupta MK, Lux MW.
  TITLE     Silk fibroin as an additive for cell-free protein synthesis
  JOURNAL   Synth Syst Biotechnol 5 (3), 145-154 (2020)
  PUBMED    32637668
REFERENCE   2  (bases 1 to 5295)
  AUTHORS   Lee MS.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-APR-2020) US Army CCDC Chemical Biological Center, 
            8567 Ricketts Point Road, Gunpowder, MD 21010, USA
REFERENCE   3  (bases 1 to 5295)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Synth Syst 
            Biotechnol"; date: "2020"; volume: "5"; issue: "3"; pages: "145-154"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-APR-2020) US Army CCDC Chemical Biological Center, 8567 Ricketts
            Point Road, Gunpowder, MD 21010, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..546)
                     /direction=LEFT
                     /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."
     CDS             complement(770..1849)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(1850..1927)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     promoter        2161..2189
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    2197..2213
                     /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)."
     gene            2277..3170
                     /gene="tyr1"
                     /label=tyr1
     CDS             2277..3170
                     /codon_start=1
                     /transl_table=11
                     /gene="tyr1"
                     /product="tyrosinase"
                     /label=tyr1
                     /note="Tyr1 tyrosinase; derived from Bacillus megaterium"
                     /protein_id="QLI61466.1"
                     /translation="MGNKYRVRKNVLHLTDTEKRDFVRTVLILKEKGIYDRYIAWHGAA
                     GKFHTPPGSDRNAAHMSSAFLPWHREYLLRFERDLQSINPEVTLPYWEWETDAQMQDPS
                     QSQIWSADFMGGNGNPIKDFIVDTGPFAAGRWTTIDEQGNPSGGLKRNFGATKEAPTLP
                     TRDDVLNALKITQYDTPPWDMTSQNSFRNQLEGFINGPQLHNRVHRWVGGQMGVVPTAP
                     NDPVFFLHHANVDRIWAVWQIIHRNQNYQPMKNGPFGQNFRDPMYPWNTTPEDVMNHRK
                     LGYVYDIELRKSKRSS"
     oriT            3587..3696
                     /label=oriT
                     /note="incP origin of transfer"
     gene            complement(4103..4741)
                     /gene="cat"
                     /label=cat
     CDS             complement(4103..4741)
                     /codon_start=1
                     /transl_table=11
                     /gene="cat"
                     /product="Cat"
                     /label=cat
                     /note="chloramphenicol acetyltransferase"
                     /protein_id="QLI61467.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRCLMNTTVLR"
     promoter        complement(4742..4844)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
ORIGIN
        1 tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg
       61 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg
      121 ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc
      181 tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc
      241 acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
      301 acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag
      361 ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc
      421 gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa
      481 ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga
      541 tctcaagaag atcatcttat taatcagata atatattttt agcggagtga ctataaaaag
      601 gcgcactgat caatgcgcct tttttataag atcataaaag acccttcatt tatgaagggt
      661 ctttttgcat gcggcgcctg atctgtcgag ccgaacgcca gcaagacgta gcccagcgcg
      721 tcggccagct tgcaattcgc gctaacttac attaattgcg ttgcgctcac tgcccgcttt
      781 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
      841 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
      901 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
      961 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
     1021 cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa
     1081 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
     1141 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
     1201 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
     1261 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
     1321 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
     1381 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
     1441 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
     1501 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
     1561 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
     1621 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
     1681 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
     1741 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
     1801 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     1861 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg caccattcga
     1921 tggtgtcaac gtaaatgcat gccgcttcgc cttcgcgcgc gaattgcaag ctgatccggg
     1981 cttatcgact gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg aagctgtggt
     2041 atggctgtgc aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc actcccgttc
     2101 tggataatgt tttttgcgcc gacatcataa cggttctggc aaatattctg aaatgagctg
     2161 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
     2221 caggaaacag cctcgacagg cctaggaatt cggcgtaagc ttaaggagat atacatatgg
     2281 gtaacaagta tagagttaga aaaaacgtat tacatcttac cgacacggaa aaaagagatt
     2341 ttgttcgtac cgtgctaata ctaaaggaaa aagggatata tgaccgctat atagcctggc
     2401 atggtgcagc aggtaaattt catactcctc cgggcagcga tcgaaatgca gcacatatga
     2461 gttctgcttt tttaccgtgg catcgtgaat accttttacg attcgaacgt gaccttcagt
     2521 caatcaatcc agaagtaacc cttccttatt gggaatggga aacggacgca cagatgcagg
     2581 atccctcaca atcacaaatt tggagtgcag attttatggg aggaaacgga aatcccataa
     2641 aagattttat cgtcgatacc gggccatttg cagctgggcg ctggacgacg atcgatgaac
     2701 aaggaaatcc ttccggaggg ctaaaacgta attttggagc aacgaaagag gcacctacac
     2761 tccctactcg agatgatgtc ctcaatgctt taaaaataac tcagtatgat acgccgcctt
     2821 gggatatgac cagccaaaac agctttcgta atcagcttga aggatttatt aacgggccac
     2881 agcttcacaa tcgcgtacac cgttgggttg gcggacagat gggcgttgtg cctactgctc
     2941 cgaatgatcc tgtcttcttt ttacaccacg caaatgtgga tcgtatttgg gctgtatggc
     3001 aaattattca tcgtaatcaa aactatcagc cgatgaaaaa cgggccattt ggtcaaaact
     3061 ttagagatcc gatgtaccct tggaatacaa cccctgaaga cgttatgaac catcgaaagc
     3121 ttgggtacgt atacgatata gaattaagaa aatcaaaacg ttcctcatga ggatccggtg
     3181 attgattgag caagctttat gcttgtaaac cgttttgtga aaaaattttt aaaataaaaa
     3241 aggggacctc tagggtcccc aattactagc caattcgatt actggcttac tatgttggca
     3301 ctgatgaggg tgtcagtgaa gtgacctgca gggggggggg gcgctgaggt ctgcctcgtg
     3361 aagaaggtgt tgctgactca taccaggctt aattcctgca tttgcctgtt tccagtcggt
     3421 agatattcca caaaacagca gggaagcagc gcttttccgc tgcataaccc tgcttcgggg
     3481 tcattatagc gattttttcg gtatatccat cctttttcgc acgatataca ggattttgcc
     3541 aaagggttcg tgtagacttt ccttggtgta tccaacggcg tcagccgggc aggataggtg
     3601 aagtaggccc acccgcgagc gggtgttcct tcttcactgt cccttattcg cacctggcgg
     3661 tgctcaacgg gaatcctgct ctgcgaggct ggccggctac cgccggcgta acagatgagg
     3721 gcaagcggat ggctgatgaa accaagccaa ccaggaaggg cagcccacct atcaaggtgt
     3781 actgccttcc agaattaagc ctgaatcgcc ccatcatcca gccagaaagt gagggagcca
     3841 cggttgatga gagctttgtt gtaggtggac cagttggtga ttttgaactt ttgctttgcc
     3901 acggaacggt ctgcgttgtc gggaagatgc gtgatctgat ccttcaactc agcaaaagtt
     3961 cgatttattc aacaaagccg ccgtcccgtc aagtcagcgt aatgctctgc cagtgttaca
     4021 accaattaac caattctaga gaataggaac ttcggaatag gaacttcatt taaatggcgc
     4081 gccttacgcc ccgccctgcc actcatcgca gtactgttgt attcattaag catctgccga
     4141 catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt
     4201 cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg
     4261 ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat
     4321 tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg
     4381 aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg
     4441 tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct
     4501 caccgtcttt cattgccata cgtaattccg gatgagcatt catcaggcgg gcaagaatgt
     4561 gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa
     4621 tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat
     4681 gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca
     4741 ttttagcttc cttagctcct gaaaatctcg acaactcaaa aaatacgccc ggtagtgatc
     4801 ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaacgtct cattttcgcc
     4861 aaaagttggc ccagggcttc ccggtatcaa cagggacacc aggatttatt tattctgcga
     4921 agtgatcttc cgtcacaggt aggcgcgccg aagttcctat actttctaga attactgttt
     4981 atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat
     5041 cagagatttt gagacacaac gtggctttcc cccccccccc tgcaggtcct tcatgtggca
     5101 ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata tattccgctt
     5161 cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg
     5221 aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa gtgagagggc
     5281 cgcggcaaag ccgtt
//
