LOCUS       sequence-319356-        3497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-term His-SNAP 3C cleavable Amp resistant bacterial expression 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3497)
  AUTHORS   Altmannova V, Blaha A, Astrinidis S, Reichle H, Weir JR
  TITLE     InteBac - An integrated bacterial and baculovirus expression vector 
            suite.
  JOURNAL   Protein Sci. 2020 Sep 21. doi: 10.1002/pro.3957.
  PUBMED    32955754
REFERENCE   2  (bases 1 to 3497)
  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: "Protein 
            Sci. 2020 Sep 21. doi: 10.1002/pro.3957."
COMMENT     Sequence Label: 17 pCOLI_A_SNAP
FEATURES             Location/Qualifiers
     source          1..3497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      95..134
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      165..753
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     654..673
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     907..924
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    939..1078
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1004..1023)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1164..1186
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1180..1371)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1608..1627
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1608..1626
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1664..1686
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1706..1723
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1742..2287
                     /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             2294..2317
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2510..2528)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2524..2571
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2636..2727
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agttatctac acgacgggta gtcaggcaac tatggacgaa cgaaatagac agattgctga
       61 aataggggct tcactgatta agcattggta aaccgataca attaaaggct ccttttggag
      121 cctttttttt tggacggacc ggtagaaaag atcaaaggat cttcttgaga tccttttttt
      181 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
      241 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
      301 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
      361 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
      421 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
      481 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
      541 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
      601 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
      661 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
      721 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
      781 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
      841 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
      901 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
      961 acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg
     1021 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     1081 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     1141 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     1201 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     1261 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     1321 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     1381 cgtgtaaggg ggatttctgt tcatggggta atgataccga tgaaacgaga gaggatgctc
     1441 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
     1501 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc
     1561 gttaatacag atgtaggtgt tggcaccatg catcgatccc gcgaaattaa tacgactcac
     1621 tatagggaga ccacaacggt ttccctccgg ctccggaaat aattttgttt aactttaaga
     1681 aggagatata catatgggca gcagccatca tcatcatcat cacagcagcg gatctggatc
     1741 tatggataaa gattgtgaaa tgaaacgtac caccctggat agcccgctgg gcaaactgga
     1801 actgagcggc tgcgaacagg gcctgcatga aattaaactg ctgggtaaag gcaccagcgc
     1861 ggccgatgcg gttgaagttc cggccccggc cgccgtgctg ggtggtccgg aaccgctgat
     1921 gcaggcgacc gcgtggctga acgcgtattt tcatcagccg gaagcgattg aagaatttcc
     1981 ggttccggcg ctgcatcatc cggtgtttca gcaggagagc tttacccgtc aggtgctgtg
     2041 gaaactgctg aaagtggtta aatttggcga agtgattagc tatcagcagc tggcggccct
     2101 ggcgggtaat ccggcggcca ccgccgccgt taaaaccgcg ctgagcggta acccggtgcc
     2161 gattctgatt ccgtgccatc gtgtggttag ctctagcggt gcggttggcg gttatgaagg
     2221 tggtctggcg gtgaaagagt ggctgctggc ccatgaaggt catcgtctgg gtaaaccggg
     2281 tctgggtgga tccctggaag ttctgttcca ggggcccgga tcccggtccg acatatgagc
     2341 gcgcggaatt caaaggccta cgtcgacgag ctcactagtc gcggccgctt tcgaatctag
     2401 agcctgcagt ctcgacaagc ttgtcgagaa gtactagagg atgtacaacg cgtgcgcgct
     2461 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     2521 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
     2581 ggaactatat cgggattagc ccgcctaatg agcgggcttt tttttaattc ccctatttgt
     2641 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2701 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     2761 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct cgtgaaagta
     2821 aaagacgcag aggaccaatt gggggcacga gtgggataca tagaactgga cttgaatagc
     2881 ggtaaaatcc ttgagagttt tcgccctgaa gagcgttttc caatgatgag cactttcaaa
     2941 gttctgctat gtggagcagt attatcccgt gtagatgcgg ggcaagagca actcggacga
     3001 cgaatacact attcgcagaa tgacttggtt gaatactccc cagtgacaga aaagcacctt
     3061 acggacggaa tgacggtaag agaattatgt agtgccgcca taacgatgag tgataacact
     3121 gcggcgaact tacttctgac aaccatcggt ggaccgaagg aattaaccgc ttttttgcac
     3181 aatatgggag accatgtaac tcgccttgac cgttgggaac cagaactgaa tgaagccata
     3241 ccaaacgacg agcgagacac cacaatgcct gcggcaatgg caacaacatt acgcaaacta
     3301 ttaactggcg aactacttac tctggcttca cggcaacaat taatagactg gcttgaagcg
     3361 gataaagttg caggaccact actgcgttcg gcacttcctg ctggctggtt tattgctgat
     3421 aaatctgggg caggagagcg tggttcacgg ggtatcattg ccgcacttgg accagatggt
     3481 aagccttccc gtatcgt
//
