LOCUS       sequence-316528-        5624 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-term 3C cleavable SNAP tag for insect cell expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5624)
  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 5624)
  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: 58 pLIB_SNAP
FEATURES             Location/Qualifiers
     source          1..5624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(23..56)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        104..195
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     120..137
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             250..279
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             253..282
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             256..285
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             304..849
                     /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             856..879
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    1108..1242
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1145..1164)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1199..1218
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1352..1517
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      1701..2155
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1788..1807)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1997..2018
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2181..2285
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2286..2354,2355..3146)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2504..2523)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3317..3905
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3806..3825
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4059..4076
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4210..4434
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4501..5034)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSIGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5223..5228,5229..5245,5246..5251))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 tcctctacgc ggccgcggta ccataacttc gtatagcata cattatacga agttatctgg
       61 ttttactagt atcgattcgc gacctactcc ggaatattaa tagatcatgg agataattaa
      121 aatgataacc atctcgcaaa taaataagta ttttactgtt ttcgtaacag ttttgtaata
      181 aaaaaaccta taaatattcc ggattattca taccgtccca ccatcgggcg cggatctatg
      241 ggcagcagcc atcatcatca tcatcaccat catcatcatc atcacagcag cggatctgga
      301 tctatggata aagattgtga aatgaaacgt accaccctgg atagcccgct gggcaaactg
      361 gaactgagcg gctgcgaaca gggcctgcat gaaattaaac tgctgggtaa aggcaccagc
      421 gcggccgatg cggttgaagt tccggccccg gccgccgtgc tgggtggtcc ggaaccgctg
      481 atgcaggcga ccgcgtggct gaacgcgtat tttcatcagc cggaagcgat tgaagaattt
      541 ccggttccgg cgctgcatca tccggtgttt cagcaggaga gctttacccg tcaggtgctg
      601 tggaaactgc tgaaagtggt taaatttggc gaagtgatta gctatcagca gctggcggcc
      661 ctggcgggta atccggcggc caccgccgcc gttaaaaccg cgctgagcgg taacccggtg
      721 ccgattctga ttccgtgcca tcgtgtggtt agctctagcg gtgcggttgg cggttatgaa
      781 ggtggtctgg cggtgaaaga gtggctgctg gcccatgaag gtcatcgtct gggtaaaccg
      841 ggtctgggtg gatccctgga agttctgttc caggggcccg gatcccggtc cgacatatga
      901 gcgcgcggaa ttcaaaggcc tacgtcgacg agctcactag tcgcggccgc tttcgaatct
      961 agagcctgca gtctcgacaa gcttgtcgag aagtactaga ggatcataat cagccatacc
     1021 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     1081 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     1141 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     1201 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tctgatcact gcttgagcct
     1261 agaagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     1321 gcaataacta tcataacccc taggagatcc gaaccagata agtgaaatct agttccaaac
     1381 tattttgtca tttttaattt tcgtattagc ttacgacgct acacccagtt cccatctatt
     1441 ttgtcactct tccctaaata atccttaaaa actccatttc cacccctccc agttcccaac
     1501 tattttgtcc gcccacagcg gggcattttt cttcctgtta tgtttttaat caaacatcct
     1561 gccaactcca tgtgacaaac cgtcatcttc ggctactttt tctctgtcac agaatgaaaa
     1621 tttttctgtc atctcttcgt tattaatgtt tgtaattgac tgaatatcaa cgcttatttg
     1681 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     1741 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     1801 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     1861 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     1921 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     1981 gtccacgttc ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2041 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2101 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     2161 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2221 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2281 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2341 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2401 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2461 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2521 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2581 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2641 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     2701 ccatcggtgg accgaaggaa ttaaccgctt ttttgcacaa catgggggat catgtaactc
     2761 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     2821 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     2881 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     2941 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3001 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3061 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3121 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3181 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3241 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3301 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3361 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3421 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tattcttcta gtgtagccgt
     3481 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3541 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3601 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3661 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3721 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3781 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3841 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3901 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3961 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4021 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     4081 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4141 tagaccagcc gcgtaacctg gcaaaatcgg ttacggttga gtaataaatg gatgccctgc
     4201 gtaagcgggt gtgggcggac aataaagtct taaactgaac aaaatagatc taaactatga
     4261 caataaagtc ttaaactaga cagaatagtt gtaaactgaa atcagtccag ttatgctgtg
     4321 aaaaagcata ctggactttt gttatggcta aagcaaactc ttcattttct gaagtgcaaa
     4381 ttgcccgtcg tattaaagag gggcgtggcc aagggcatgg taaagactat attcgcggcg
     4441 ttgtgacaat ttaccgaaca actccgcggc cgggaagccg atctcggctt gaacgaattg
     4501 ttaggtggcg gtacttgggt cgatatcaaa gtgcatcact tcttcccgta tgcccaactt
     4561 tgtatagaga gccactgcgg gatcgtcacc gtaatctgct tgcacgtaga tcacataagc
     4621 accaagcgcg ttggcctcat gcttgaggag attgatgagc gcggtggcaa tgccctgcct
     4681 ccggtgctcg ccggagactg cgagatcata gatatagatc tcactacgcg gctgctcaaa
     4741 cttgggcaga acgtaagccg cgagagcgcc aacaaccgct tcttggtcga aggcagcaag
     4801 cgcgatgaat gtcttactac ggagcaagtt cccgaggtaa tcggagtccg gctgatgttg
     4861 ggagtaggtg gctacgtctc cgaactcacg accgaaaaga tcaagagcag cccgcatgga
     4921 tttgacttgg tcagggccga gcctacatgt gcgaatgatg ccaatacttg agccacctaa
     4981 ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctgcgta acatcgttgc
     5041 tgctccataa catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag
     5101 gcatagactg tacaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac
     5161 caccgctgcg ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt
     5221 acagtttacg aaccgaacag gcttatgtca actgggtttg tgccttcatc cgtttccacg
     5281 gtgtgcgtca cccggcaacc ttgggcagca gcgaagtcga ggcatttctg tcctggctgg
     5341 cgaacgagcg caaggtttcg gtctccacgc atcgtcaggc attggcggcc ttgctgttct
     5401 tctacggcaa ggtgctgtgc acggatctgc ccttgcttca ggagatcggt agacctcggc
     5461 cgtcgcggcg cttgccggtg gtgctgaccc cggatgaagt ggttcgcatc ctcggttttc
     5521 tggaaggcga gcatcgtttg ttcgcccagg actctagcta tagttctagt ggttggctac
     5581 agctttgttt gtactatcaa caggttgaac tgctgatcaa caga
//
