LOCUS       sequence-142447-        6414 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Photinus pyralis sulfotransferase ST3 - pHis8-4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6414)
  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 6414)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Biochemistry. 2016 May 26."
COMMENT     Sequence Label: pJKW 0690
FEATURES             Location/Qualifiers
     source          1..6414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             4175..4192
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4245..4263)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4259..4306
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4343..4798
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4430..4449)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4640..4661
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4891..5706)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4891..5706)
                     /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     5614..5633
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5698..5717
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(5828..6414,1..2)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6317..6336
                     /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 atgccacata
     3061 acattcaaat tggggatatt ttaaataata gagatgaaaa tagtaagagg aaatttcctt
     3121 acgaagttaa aattgtagat cctgacacaa ataaggagct attgaatgtt tttacagggg
     3181 agcatacagg gttcgtacaa gtaggccctc aaaaatattt ctttcctagc ggctatgaaa
     3241 aggtagcaga aaaatattac aattttgaaa tacgacccga cgatgtctgg attgttacat
     3301 ttccaagatc aggtacaacg tggacgcaag aaatggtgtg gttaattgca aacgaatttg
     3361 actacaatac ggccgctaga gtaccattaa acgaaagatt tcctttttta gaattcagca
     3421 tgtgcgtcca tccgaaaatg aaggcacaat tcttacaaga aaatagtcac agtattgaac
     3481 gacaaaatat gatcgaagac gtctgcacag aagcatggaa gattttggaa aattggaagg
     3541 ggaggcgttt catcaaaaca cacctgccat tcagcctgtt gccacctaat ttgttaacct
     3601 ctggctgtaa ggtaatttac gttgcaagaa acccgaagga tgttatggtc tcgtactact
     3661 atttgaacag attatttcga acccaagggt atcggggtga ctttccgaca ttcgcggagt
     3721 acttcctaaa ggatatggta cattggacgc cgttttggtc tcatttacaa gaaggctggg
     3781 atttgcgaag ctccaagaat cttctttttc tgttctacga ccaaatgaat aaattgcaga
     3841 caatccaaaa ggtgtcgaac tttttgggga aggaaattga tgacgatcaa ttaacgaaaa
     3901 tacagaatca tttacaaatt gagaacttta agaataataa gtcgctcaac gctaatgatt
     3961 tattagaatt gggaatatta tatggaggcg aagagggttt tgtaaggaag ggaaagacgg
     4021 gatcacatac ggacttatcc aatgtcgaat tagacgagaa gattgacaag tgggtcaaaa
     4081 agaatttaca gatgacgaat ataccgtttg aacgaatgta accatggcgg atccgaattc
     4141 gagctccgtc gacaagcttg cggccgcact cgagcaccac caccaccacc actgagatcc
     4201 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     4261 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     4321 tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4381 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4441 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4501 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4561 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4621 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4681 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4741 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttca
     4801 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4861 tcaaatatgt atccgctcat gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa
     4921 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     4981 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     5041 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     5101 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg
     5161 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     5221 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct
     5281 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     5341 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc
     5401 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     5461 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     5521 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     5581 tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     5641 atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg
     5701 gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgacca
     5761 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5821 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5881 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5941 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     6001 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6061 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6121 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6181 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6241 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6301 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6361 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tgga
//
