LOCUS       Exported                6219 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Photinus pyralis sulfotransferase ST2 - pHis8-4.
ACCESSION   .
VERSION     .
KEYWORDS    pJKW 0633
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6219)
  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 6219)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6219
                     /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             3980..3997
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4050..4068)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4064..4111
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4148..4603
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4235..4254)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4445..4466
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4696..5511)
                     /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     5419..5438
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5503..5522
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5633..2
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6122..6141
                     /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 atggaagaaa
     3061 ataactatct cccttatttg tgtgaaaaaa ccgcaaggag tctttgcgcc aaaatcaaac
     3121 cgtttgaagt taaaaatgac gacatttttg taattggagc gacaagatcg ggtaccatgt
     3181 gggtgcagga aatggtttgg ttgattgcaa atgatttgga ttatgagaaa gctaaagagg
     3241 gggcgtacta ttgggttcca cttctggaat atagtgttta tttgtgcagc aaaccgggct
     3301 ctcagtctta tcatatgcaa gagcggtacg agcctaattc aatagagttt tgtaaaaact
     3361 taaaaagtcc acgcataatt aaaacgcatt taccagaaga atatctacca cggcaactgt
     3421 ttgataaaga aaaggaaact aagattattt acattgcacg caaccccaag gatgtctgct
     3481 tgtcaggata tcattttaaa agaattgtta tgaagtgcga atttgaatcc ctggagagct
     3541 atgccgaagt tttcatgtcg gattcatttc ccaaaggaga ttattggaag aatgtgctgt
     3601 acttttggaa ccgtcgccac gaggataata ttttgtttat tacgtacgaa gaaatgaaga
     3661 aagatttgag ggcggttatg cggaaagtgg ccgaatttct tggcaaagtt ttgacgatcg
     3721 aacaggagga ggagttgcaa agatacctgg actttgggtc gttcaaaaac aacaatgccc
     3781 tcaatcaaat tgactttttt aatgcagatg catatttaag acgaggggta gttggaggat
     3841 acaaaacgga aatgtccaat gaattgatcg ataagtttga tagttggagc aaggaatgtc
     3901 ttaaacattc tgattataaa ttataaccat ggcggatccg aattcgagct ccgtcgacaa
     3961 gcttgcggcc gcactcgagc accaccacca ccaccactga gatccggctg ctaacaaagc
     4021 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     4081 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc
     4141 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4201 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4261 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4321 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4381 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4441 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4501 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4561 aaatttaacg cgaattttaa caaaatatta acgtttacaa tttcaggtgg cacttttcgg
     4621 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4681 ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     4741 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     4801 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     4861 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     4921 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
     4981 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     5041 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     5101 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     5161 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     5221 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     5281 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     5341 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     5401 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     5461 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct
     5521 tgtattactg tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg
     5581 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5641 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5701 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5761 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5821 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5881 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5941 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6001 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6061 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6121 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6181 gatttttgtg atgctcgtca ggggggcgga gcctatgga
//
