LOCUS       sequence-315214-        5670 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid for AcpS phosphopantetheinyl transferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5670)
  AUTHORS   Zhou Z, Cironi P, Lin AJ, Xu Y, Hrvatin S, Golan DE, Silver PA, 
            Walsh CT, Yin J
  TITLE     Genetically encoded short peptide tags for orthogonal protein 
            labeling by Sfp and AcpS phosphopantetheinyl transferases.
  JOURNAL   ACS Chem Biol. 2007 May 22;2(5):337-46. doi: 10.1021/cb700054k. Epub
            2007 Apr 27.
  PUBMED    17465518
REFERENCE   2  (bases 1 to 5670)
  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"; doi: "10.1021/cb700054k";
            journalName: "ACS Chem Biol"; date: "2007-05-22- 22"; volume: "2"; 
            issue: "5"; pages: "337-46"
COMMENT     Sequence Label: pET-AspS
FEATURES             Location/Qualifiers
     source          1..5670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             15..37
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             56..73
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             83..100
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             491..508
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(561..579)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      575..622
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      659..1114
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(746..765)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     956..977
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1206..2021)
                     /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(1206..2021)
                     /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     1929..1948
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2013..2032
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2143..2731
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2632..2651
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2885..2902
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2917..3056
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2982..3001)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3142..3164
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3158..3349)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4158..5240)
                     /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     5202..5221
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5241..5318)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5524..5543
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5627..5646
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5627..5645
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5646..5670
                     /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)."
ORIGIN
        1 cctctagaaa taattttgtt taactttaag aaggagatat accatgggca gcagccatca
       61 tcatcatcat cacagcagcg gcctggtgcc gcgcggcagc catatggcaa tattaggttt
      121 aggcacggat attgtggaga tcgctcgcat cgaagcggtg atcgcccgat ccggtgatcg
      181 cctggcacgc cgcgtattaa gcgataacga atgggctatc tggaaaacgc accaccagcc
      241 ggtgcgtttt ctggcgaagc gttttgctgt gaaagaagcc gcagcaaaag cgtttggcac
      301 cgggatccgc aatggtctgg cgtttaatca atttgaagta ttcaatgatg agctcggcaa
      361 accacggcta cggctatggg gcgaggcatt aaaactggcg gaaaagctgg gcgttgcaaa
      421 tatgcatgta acgctggcag atgagcggca ctatgcttgt gccacggtaa ttattgaaag
      481 ttaactcgag caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga
      541 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
      601 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac
      661 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
      721 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
      781 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
      841 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
      901 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
      961 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1021 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1081 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     1141 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgaatt
     1201 aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     1261 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     1321 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     1381 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     1441 acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca
     1501 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     1561 gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     1621 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     1681 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     1741 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     1801 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     1861 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     1921 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     1981 cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
     2041 tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg agttttcgtt
     2101 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2161 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2221 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2281 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2341 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2401 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2461 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2521 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2581 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2641 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2701 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2761 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     2821 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     2881 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     2941 gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc
     3001 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     3061 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     3121 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     3181 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     3241 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     3301 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     3361 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     3421 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     3481 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     3541 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     3601 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     3661 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     3721 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     3781 tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg
     3841 cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt
     3901 gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt
     3961 cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga
     4021 agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg
     4081 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     4141 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4201 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     4261 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     4321 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     4381 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata
     4441 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     4501 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     4561 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     4621 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     4681 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     4741 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     4801 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     4861 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     4921 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     4981 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     5041 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     5101 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     5161 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     5221 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     5281 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     5341 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     5401 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     5461 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     5521 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     5581 cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact
     5641 ataggggaat tgtgagcgga taacaattcc
//
