LOCUS       Exported                6456 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Photinus pyralis luciferin sulfotransferase LST - pHis8-4.
ACCESSION   .
VERSION     .
KEYWORDS    pJKW 0643
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6456)
  AUTHORS   Fallon TR, Li FS, Vicent MA, Weng JK
  TITLE     Sulfoluciferin is biosynthesized by a specialized luciferin 
            sulfotransferase in fireflies.
  JOURNAL   Biochemistry. 2016 May 26.
  PUBMED    27227579
REFERENCE   2  (bases 1 to 6456)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6456
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     156..173
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    188..330
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(256..275)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     416..438
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(432..623)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1432..2514)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2476..2495
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2515..2592)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2798..2817
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2901..2920
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2901..2919
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2920..2944
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     RBS             2959..2981
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2994..3017
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             3024..3044
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQA"
     CDS             4217..4234
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4287..4305)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4301..4348
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4385..4840
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4472..4491)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4682..4703
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4933..5748)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5656..5675
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5740..5759
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5870..2
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6359..6378
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
       61 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      121 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      181 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
      241 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
      301 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
      361 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
      421 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
      481 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
      541 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
      601 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
      661 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
      721 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
      781 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
      841 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
      901 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
      961 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     1021 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     1081 cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
     1141 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
     1201 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
     1261 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     1321 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     1381 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     1441 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     1501 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     1561 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     1621 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     1681 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     1741 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     1801 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     1861 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     1921 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     1981 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     2041 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     2101 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     2161 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     2221 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     2281 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     2341 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     2401 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     2461 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     2521 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     2581 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     2641 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     2701 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     2761 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     2821 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
     2881 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
     2941 ttcccctcta gaaataattt tgtttaactt taagaaggag atataccatg aaacaccacc
     3001 accaccacca ccaccacggt ggtgaaaact tgtacttcca ggcccatggc atgtttgcat
     3061 ctatcctagg caaaatacca caatattcta cgatttcgtc aaaatgttcg ttaaaagaat
     3121 gtcgaaaagt taagaaagta gagaactctg atgaaatcgg cgaaattatt catcgggact
     3181 ttttagatcc gttttgtaat gagtatgtat taattggtga ggaagagacc tctttaggta
     3241 tcaattattt gcaatttgaa gaagaaatta gaactttcga aattcgcgat tcagatatta
     3301 ttgtcgcttc atatcccaag gcaggcacta catggaccca ggagttggtg tggttaattg
     3361 gaaatgattt agactttaaa gcggcagaag aacacttaga caaacgattt cctcatttcg
     3421 aattatgcac catagtaaat tttgctaaaa tgaccgaaat gttaggatct acacgtccag
     3481 aatacatagg taattcaatt aactatttga gagatttaga agggactcgt ttcattaaga
     3541 cacatttaac atataactta ttaccagagc aaattcttaa tgggaataga aaaccaaaga
     3601 ttatatatgt catgcgggat ccaaaagacg tatgcgtgtc ttattaccat cacggtaggc
     3661 tcatacaagg gtggcgtgcc gactttcaaa acttttccaa agtatttctc tcagaaaaaa
     3721 ttatgtttgg ctcttattgg aagcacgtat taggatattg ggaacatcga gacaaaccta
     3781 atgtgttaat acttacatat gaagagatga aaaaggatct tctatctgtt ataaggaaaa
     3841 ctgcccaatt tttagataag aaattgaacg aaaacaaaat accgcaactg ttgaagcact
     3901 taagtttcga atctatgaaa aacaatcgag ccgttaatca gcaagataaa attgaaagta
     3961 gaatgaaaca caagttggtg ccagagcaag gtgcatttat gaggtccggc acttctcaaa
     4021 attataaagg ggaaatgtct gaagaattga ttttaaaatt tgacgaatgg acaaaaaaat
     4081 caatctctgg cagttcattc gtaaccgagc ccatagcaga cttatatctg aatacttctc
     4141 aaaaatctgt tccaaaaatg taaccatggc ggatccgaat tcgagctccg tcgacaagct
     4201 tgcggccgca ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg
     4261 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     4321 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa
     4381 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4441 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4501 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4561 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4621 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4681 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4741 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4801 tttaacgcga attttaacaa aatattaacg tttacaattt caggtggcac ttttcgggga
     4861 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4921 atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
     4981 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     5041 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     5101 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     5161 atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg
     5221 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
     5281 cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca
     5341 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     5401 tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag
     5461 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     5521 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     5581 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc
     5641 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     5701 atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa caccccttgt
     5761 attactgttt atgtaagcag acagttttat tgttcatgac caaaatccct taacgtgagt
     5821 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5881 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5941 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6001 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6061 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6121 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6181 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6241 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6301 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6361 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6421 ttttgtgatg ctcgtcaggg gggcggagcc tatgga
//
