LOCUS       sequence-493095-        6297 bp    DNA     circular SYN 18-NOV-2025
DEFINITION  Protein Transposon Cassette Containing CatC and CfaCNm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6297)
  TITLE     40179182
REFERENCE   2  (bases 1 to 6297)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 18, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pET-CatC-HA-CfaNm
FEATURES             Location/Qualifiers
     source          1..6297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             117..134
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(187..205)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      201..248
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      285..740
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(372..391)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     582..603
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(832..1647)
                     /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(832..1647)
                     /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     1555..1574
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1639..1658
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1769..2357
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2258..2277
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2511..2528
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2543..2682
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2608..2627)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2768..2790
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2784..2975)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3784..4866)
                     /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     4828..4847
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4867..4944)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5150..5169
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5257..5276
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5257..5275
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5276..5300
                     /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             5315..5337
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5381..5398
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5405..5698
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             6029..6055
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 cgacggtcag ctgttaccga ttgatgagat tttcgaacgg gggttagacc tgaaacaagt
       61 tgatggtttg ccgtaagaat tcgagctccg tcgacaagct tgcggccgca ctcgagcacc
      121 accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg
      181 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
      241 gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg
      301 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
      361 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
      421 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
      481 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
      541 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
      601 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
      661 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
      721 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
      781 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa
      841 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
      901 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
      961 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     1021 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     1081 tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     1141 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     1201 gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     1261 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     1321 tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt
     1381 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     1441 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     1501 cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg
     1561 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca
     1621 agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga
     1681 cagttttatt gttcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     1741 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1801 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1861 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     1921 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1981 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2041 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2101 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2161 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2221 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2281 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2341 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     2401 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     2461 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     2521 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     2581 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     2641 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     2701 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     2761 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     2821 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     2881 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     2941 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     3001 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     3061 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     3121 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     3181 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     3241 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     3301 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     3361 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     3421 agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg
     3481 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     3541 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     3601 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     3661 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     3721 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     3781 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     3841 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     3901 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     3961 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     4021 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg
     4081 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     4141 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     4201 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     4261 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     4321 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     4381 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     4441 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     4501 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     4561 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     4621 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     4681 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     4741 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     4801 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     4861 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     4921 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
     4981 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccaca aggaatggtg
     5041 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     5101 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     5161 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     5221 cgtagaggat cgagatcgat ctcgatcccg cgaaattaat acgactcact ataggggaat
     5281 tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat
     5341 acatatgggc agcagccatc atcatcatca cggcagcggc ctggtgccgc gcggcagcgc
     5401 tagcatgtcg gactcagaag tcaatcaaga agctaagcca gaggtcaagc cagaagtcaa
     5461 gcctgagact cacatcaatt taaaggtgtc cgatggatct tcagagatct tcttcaagat
     5521 caaaaagacc actcctttaa gaaggctgat ggaagcgttc gctaaaagac agggtaagga
     5581 aatggactcc ttaagattct tgtacgacgg tattagaatt caagctgatc agacccctga
     5641 agatttggac atggaggata acgatattat tgaggctcac agagaacaga ttggtggtat
     5701 gttcaaattg aacacgaaaa acatcaaggt tttaacgccg tctggattca aatctttctc
     5761 tggtatccag aaagtctata aaccgttcta ccaccatatc atttttgacg acgggtcgga
     5821 gatcaagtgt tctgacaatc attcctttgg aaaggataaa atcaaagcgt cgacgatcaa
     5881 agtgggcgac tatttacagg gaaaaaaagt actttataat gagatcgtag aggaaggaat
     5941 ttatctgtac gacctgctta acgtggggga agataactta tattacacta atggaatcgt
     6001 cagccataac tgcgaagcct cggggaagta cccttacgac gttcctgact acgctggcgc
     6061 cgagtattgc ctgtcttacg acacagagat tctgaccgtt gaatatggat tccttcctat
     6121 cggtaagatc gtggaggaac ggattgaatg cacagtctat acggtagata aaaatggctt
     6181 tgtgtataca caacctattg ctcagtggca taaccgggga gaacaggaag ttttcgaata
     6241 ctgcttagaa gacggttcga ttatccgtgc aacgaaagat cacaaatttc tgacgac
//
