LOCUS       sequence-211598-        5678 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression plasmid for C-terminal his tagged, calcium independent 
            S.aureus SrtA with enhanced catalytic activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5678)
  TITLE     unpublished
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5678)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pet30b-7M SrtA
FEATURES             Location/Qualifiers
     source          1..5678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     489..508
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     742..759
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    774..913
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(839..858)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     999..1021
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1015..1206)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2015..3097)
                     /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     3059..3078
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3098..3175)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3381..3400
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3488..3507
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3488..3506
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3507..3531
                     /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             3546..3568
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4026..4043
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4096..4114)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4110..4157
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4194..4649
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4281..4300)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4491..4512
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4741..5556)
                     /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(4741..5556)
                     /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     5464..5483
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5548..5567
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(5678,1..588)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
       61 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
      121 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
      181 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
      241 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
      301 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
      361 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
      421 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
      481 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
      541 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
      601 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
      661 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
      721 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
      781 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
      841 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
      901 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
      961 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     1021 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     1081 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     1141 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     1201 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     1261 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     1321 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     1381 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     1441 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     1501 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     1561 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     1621 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     1681 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     1741 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     1801 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     1861 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     1921 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     1981 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2041 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2101 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     2161 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     2221 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     2281 atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     2341 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     2401 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     2461 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     2521 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     2581 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     2641 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     2701 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     2761 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     2821 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     2881 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     2941 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     3001 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     3061 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     3121 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     3181 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     3241 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     3301 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     3361 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     3421 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatcga tctcgatccc
     3481 gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa
     3541 ataattttgt ttaactttaa gaaggagata tacatatgca agctaaacct caaattccga
     3601 aagataaatc aaaagtggca ggctatattg aaattccaga tgctgatatt aaagaaccag
     3661 tatatccagg accagcaaca cgcgaacaat taaatagagg tgtaagcttt gcaaaagaaa
     3721 atcaatcact agatgatcaa aatatttcaa ttgcaggaca cactttcatt gaccgtccga
     3781 actatcaatt tacaaatctt aaagcagcca aaaaaggtag tatggtgtac tttaaagttg
     3841 gtaatgaaac acgtaagtat aaaatgacaa gtataagaaa cgttaagcca acagctgtag
     3901 aagttctaga tgaacaaaaa ggtaaagata aacaattaac attaattact tgtgatgatt
     3961 acaatgaaga gacaggcgtt tgggaaacac gtaaaatctt tgtagctaca gaagtcaaac
     4021 tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg
     4081 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     4141 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc
     4201 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4261 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4321 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4381 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4441 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4501 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4561 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4621 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     4681 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc
     4741 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
     4801 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     4861 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     4921 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     4981 tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca
     5041 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     5101 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     5161 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     5221 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
     5281 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     5341 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     5401 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     5461 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     5521 cctagagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
     5581 gtaagcagac agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact
     5641 gagcgtcaga ccccgtagaa aagatcaaag gatcttct
//
