LOCUS       sequence-317230-        5069 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-term 3C cleavable His tag for insect cell expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5069)
  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 5069)
  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: 46 pLIB_3C_C_HIS
FEATURES             Location/Qualifiers
     source          1..5069
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      512..1100
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1001..1020
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1254..1271
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  1405..1629
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1696..2229)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2418..2423,2424..2440,2441..2446))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(2842..2875)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2923..3014
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2939..2956
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             3177..3200
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3207..3236
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             3210..3239
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             3213..3242
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     polyA_signal    3372..3506
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3409..3428)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3463..3482
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3616..3781
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3965..4419
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4052..4071)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4261..4282
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4445..4549
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4550..4618,4619..5069,1..341)
                     /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(4768..4787)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
       61 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      121 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      181 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      241 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
      301 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
      361 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
      421 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
      481 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
      541 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
      601 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactattc
      661 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
      721 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
      781 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
      841 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
      901 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
      961 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1021 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1081 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1141 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1201 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1261 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     1321 gtatttcaca ccgcatagac cagccgcgta acctggcaaa atcggttacg gttgagtaat
     1381 aaatggatgc cctgcgtaag cgggtgtggg cggacaataa agtcttaaac tgaacaaaat
     1441 agatctaaac tatgacaata aagtcttaaa ctagacagaa tagttgtaaa ctgaaatcag
     1501 tccagttatg ctgtgaaaaa gcatactgga cttttgttat ggctaaagca aactcttcat
     1561 tttctgaagt gcaaattgcc cgtcgtatta aagaggggcg tggccaaggg catggtaaag
     1621 actatattcg cggcgttgtg acaatttacc gaacaactcc gcggccggga agccgatctc
     1681 ggcttgaacg aattgttagg tggcggtact tgggtcgata tcaaagtgca tcacttcttc
     1741 ccgtatgccc aactttgtat agagagccac tgcgggatcg tcaccgtaat ctgcttgcac
     1801 gtagatcaca taagcaccaa gcgcgttggc ctcatgcttg aggagattga tgagcgcggt
     1861 ggcaatgccc tgcctccggt gctcgccgga gactgcgaga tcatagatat agatctcact
     1921 acgcggctgc tcaaacttgg gcagaacgta agccgcgaga gcgccaacaa ccgcttcttg
     1981 gtcgaaggca gcaagcgcga tgaatgtctt actacggagc aagttcccga ggtaatcgga
     2041 gtccggctga tgttgggagt aggtggctac gtctccgaac tcacgaccga aaagatcaag
     2101 agcagcccgc atggatttga cttggtcagg gccgagccta catgtgcgaa tgatgcccat
     2161 acttgagcca cctaactttg ttttagggcg actgccctgc tgcgtaacat cgttgctgct
     2221 gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa cgcgcttgct
     2281 gcttggatgc ccgaggcata gactgtacaa aaaaacagtc ataacaagcc atgaaaaccg
     2341 ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca
     2401 tacgctactt gcattacagt ttacgaaccg aacaggctta tgtcaactgg gtttgtgcct
     2461 tcatccgttt ccacggtgtg cgtcacccgg caaccttggg cagcagcgaa gtcgaggcat
     2521 ttctgtcctg gctggcgaac gagcgcaagg tttcggtctc cacgcatcgt caggcattgg
     2581 cggccttgct gttcttctac ggcaaggtgc tgtgcacgga tctgcccttg cttcaggaga
     2641 tcggtagacc tcggccgtcg cggcgcttgc cggtggtgct gaccccggat gaagtggttc
     2701 gcatcctcgg ttttctggaa ggcgagcatc gtttgttcgc ccaggactct agctatagtt
     2761 ctagtggttg gctacagctt tgtttgtact atcaacaggt tgaactgctg atcaacagat
     2821 cctctacgcg gccgcggtac cataacttcg tatagcatac attatacgaa gttatctggt
     2881 tttactagta tcgattcgcg acctactccg gaatattaat agatcatgga gataattaaa
     2941 atgataacca tctcgcaaat aaataagtat tttactgttt tcgtaacagt tttgtaataa
     3001 aaaaacctat aaatattccg gattattcat accgtcccac catcgggcgc ggatcccggt
     3061 ccgacatatg agcgcgcgga attcaaaggc ctacgtcgac gagctcacta gtcgcggccg
     3121 ctttcgaatc tagagcctgc agtctcgaca agcttggaag cggatctgga tctggactgg
     3181 aagttctgtt ccaggggccc ggtagccatc atcatcatca tcatcatcac catcaccatc
     3241 actaataagt cgagaagtac tagaggatca taatcagcca taccacattt gtagaggttt
     3301 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     3361 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3421 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3481 tcaatgtatc ttatcatgtc tggatctgat cactgcttga gcctagaaga tccggctgct
     3541 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actatcataa
     3601 cccctaggag atccgaacca gataagtgaa atctagttcc aaactatttt gtcattttta
     3661 attttcgtat tagcttacga cgctacaccc agttcccatc tattttgtca ctcttcccta
     3721 aataatcctt aaaaactcca tttccacccc tcccagttcc caactatttt gtccgcccac
     3781 agcggggcat ttttcttcct gttatgtttt taatcaaaca tcctgccaac tccatgtgac
     3841 aaaccgtcat cttcggctac tttttctctg tcacagaatg aaaatttttc tgtcatctct
     3901 tcgttattaa tgtttgtaat tgactgaata tcaacgctta tttgcagcct gaatggcgaa
     3961 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4021 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4081 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4141 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4201 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttcttaata
     4261 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4321 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4381 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     4441 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4501 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4561 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4621 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4681 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4741 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4801 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4861 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4921 cataaccatg agtgataaca ctgcggccaa cttacttctg acaaccatcg gtggaccgaa
     4981 ggaattaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5041 accggagctg aatgaagcca taccaaacg
//
