LOCUS       sequence-468942-        5728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for SNAPtag (carboxy terminal 3C-8His tag).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5728)
  TITLE     Site-specific DNA bioconjugation with polymer beads using 
            SNAP-tag(R) technology
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5728)
  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            "Unpublished"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: pSG7163 pLIBT7_SNAPtag-3C-8His
FEATURES             Location/Qualifiers
     source          1..5728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(56..75)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     216..238
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(232..423)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1232..2320)
                     /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
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQTGGGNANQLLPVSLVKRKTTLAPNTQTAS
                     PRALADSLMQLARQVSRLESGQ"
     primer_bind     2282..2301
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2321..2398)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2591..2610
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2591..2609
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2610..2634
                     /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             2649..2671
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2679..3224
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="optimized for bacterial expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     CDS             3228..3251
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3252..3275
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      3280..3327
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3415..3434)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3625..3646
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3809..3913
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3914..3982,3983..4774)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4132..4151)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4945..5533
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5434..5453
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    join(5719..5728,1..130)
                     /label=bom
                     /note="basis of mobility region from pBR322"
ORIGIN
        1 tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa
       61 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
      121 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
      181 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
      241 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
      301 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
      361 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
      421 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
      481 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
      541 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
      601 atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc
      661 gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga
      721 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
      781 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
      841 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat
      901 gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc
      961 ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct
     1021 ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag
     1081 ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg
     1141 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg
     1201 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     1261 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     1321 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg gcgttgccac
     1381 ctccagtctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc
     1441 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt
     1501 cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca
     1561 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat
     1621 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg
     1681 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg
     1741 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca
     1801 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg
     1861 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa
     1921 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa
     1981 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca
     2041 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt
     2101 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt
     2161 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg
     2221 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac
     2281 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac
     2341 tctcttccgg gcgctatcat accataccgc gaaaggtttt gcgccattcg atggtgtccg
     2401 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg
     2461 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     2521 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     2581 ctcttcccga taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta
     2641 gaaataattt tgtttaactt taagaaggag atataaccat ggacaaagat tgcgaaatga
     2701 aacgtaccac cctggatagc ccgctgggca aactggaact gagcggctgc gaacagggcc
     2761 tgcatgaaat taaactgctg ggtaaaggca ccagcgcggc cgatgcggtt gaagttccgg
     2821 ccccggccgc cgtgctgggt ggtccggaac cgctgatgca ggcgaccgcg tggctgaacg
     2881 cgtattttca tcagccggaa gcgattgaag aatttccggt tccggcgctg catcatccgg
     2941 tgtttcagca ggagagcttt acccgtcagg tgctgtggaa actgctgaaa gtggttaaat
     3001 ttggcgaagt gattagctat cagcagctgg cggccctggc gggtaatccg gcggccaccg
     3061 ccgccgttaa aaccgcgctg agcggtaacc cggtgccgat tctgattccg tgccatcgtg
     3121 tggttagctc tagcggtgcg gttggcggtt atgaaggtgg tctggcggtg aaagagtggc
     3181 tgctggccca tgaaggtcat cgtctgggta aaccgggtct gggatccctg gaagttctgt
     3241 tccaggggcc ccaccatcac catcaccatc accattaagc tagcataacc ccttggggcc
     3301 tctaaacggg tcttgagggg ttttttgggg accggaagag cggcgcatta agcgcggcgg
     3361 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3421 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3481 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3541 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3601 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3661 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3721 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     3781 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3841 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3901 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3961 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4021 tcagttgggt gcacgggtgg gttacatcga actggatctc aacagcggta agatccttga
     4081 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4141 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4201 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4261 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4321 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4381 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4441 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4501 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4561 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4621 tgagcgtggg tcccgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4681 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4741 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4801 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4861 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4921 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4981 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5041 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     5101 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5161 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5221 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5281 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5341 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5401 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5461 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5521 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5581 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5641 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     5701 agaggaagcg gatgagcgcc tgatgcgg
//
