LOCUS       sequence-317228-        5066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-term 3C cleavable 12xHis tag for insect cell expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5066)
  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 5066)
  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: 44 pLIB_12HIS
FEATURES             Location/Qualifiers
     source          1..5066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(89..122)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        170..261
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     186..203
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             316..345
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             319..348
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             322..351
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             364..387
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    616..750
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(653..672)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     707..726
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  860..1025
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      1209..1663
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1296..1315)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1505..1526
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1689..1793
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1794..1862,1863..2654)
                     /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(2012..2031)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2825..3413
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3314..3333
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3567..3584
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  3718..3942
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4009..4542)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSIGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4731..4736,4737..4753,4754..4759))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 tatagttcta gtggttggct acagctttgt ttgtactatc aacaggttga actgctgatc
       61 aacagatcct ctacgcggcc gcggtaccat aacttcgtat agcatacatt atacgaagtt
      121 atctggtttt actagtatcg attcgcgacc tactccggaa tattaataga tcatggagat
      181 aattaaaatg ataaccatct cgcaaataaa taagtatttt actgttttcg taacagtttt
      241 gtaataaaaa aacctataaa tattccggat tattcatacc gtcccaccat cgggcgcgga
      301 tctatgggca gcagccatca tcatcatcat caccatcatc atcatcatca cagcagcgga
      361 tctctggaag ttctgttcca ggggcccgga tcccggtccg acatatgagc gcgcggaatt
      421 caaaggccta cgtcgacgag ctcactagtc gcggccgctt tcgaatctag agcctgcagt
      481 ctcgacaagc ttgtcgagaa gtactagagg atcataatca gccataccac atttgtagag
      541 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
      601 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
      661 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
      721 ctcatcaatg tatcttatca tgtctggatc tgatcactgc ttgagcctag aagatccggc
      781 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactatc
      841 ataaccccta ggagatccga accagataag tgaaatctag ttccaaacta ttttgtcatt
      901 tttaattttc gtattagctt acgacgctac acccagttcc catctatttt gtcactcttc
      961 cctaaataat ccttaaaaac tccatttcca cccctcccag ttcccaacta ttttgtccgc
     1021 ccacagcggg gcatttttct tcctgttatg tttttaatca aacatcctgc caactccatg
     1081 tgacaaaccg tcatcttcgg ctactttttc tctgtcacag aatgaaaatt tttctgtcat
     1141 ctcttcgtta ttaatgtttg taattgactg aatatcaacg cttatttgca gcctgaatgg
     1201 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     1261 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     1321 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     1381 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     1441 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     1501 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     1561 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     1621 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     1681 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1741 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1801 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1861 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1921 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1981 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     2041 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2101 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2161 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacc atcggtggac
     2221 cgaaggaatt aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     2281 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     2341 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     2401 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     2461 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     2521 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     2581 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     2641 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     2701 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     2761 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2821 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2881 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2941 gcttcagcag agcgcagata ccaaatacta ttcttctagt gtagccgtag ttaggccacc
     3001 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3061 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3121 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3181 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3241 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3301 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3361 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3421 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     3481 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     3541 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     3601 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata gaccagccgc
     3661 gtaacctggc aaaatcggtt acggttgagt aataaatgga tgccctgcgt aagcgggtgt
     3721 gggcggacaa taaagtctta aactgaacaa aatagatcta aactatgaca ataaagtctt
     3781 aaactagaca gaatagttgt aaactgaaat cagtccagtt atgctgtgaa aaagcatact
     3841 ggacttttgt tatggctaaa gcaaactctt cattttctga agtgcaaatt gcccgtcgta
     3901 ttaaagaggg gcgtggccaa gggcatggta aagactatat tcgcggcgtt gtgacaattt
     3961 accgaacaac tccgcggccg ggaagccgat ctcggcttga acgaattgtt aggtggcggt
     4021 acttgggtcg atatcaaagt gcatcacttc ttcccgtatg cccaactttg tatagagagc
     4081 cactgcggga tcgtcaccgt aatctgcttg cacgtagatc acataagcac caagcgcgtt
     4141 ggcctcatgc ttgaggagat tgatgagcgc ggtggcaatg ccctgcctcc ggtgctcgcc
     4201 ggagactgcg agatcataga tatagatctc actacgcggc tgctcaaact tgggcagaac
     4261 gtaagccgcg agagcgccaa caaccgcttc ttggtcgaag gcagcaagcg cgatgaatgt
     4321 cttactacgg agcaagttcc cgaggtaatc ggagtccggc tgatgttggg agtaggtggc
     4381 tacgtctccg aactcacgac cgaaaagatc aagagcagcc cgcatggatt tgacttggtc
     4441 agggccgagc ctacatgtgc gaatgatgcc aatacttgag ccacctaact ttgttttagg
     4501 gcgactgccc tgctgcgtaa catcgttgct gctgcgtaac atcgttgctg ctccataaca
     4561 tcaaacatcg acccacggcg taacgcgctt gctgcttgga tgcccgaggc atagactgta
     4621 caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt
     4681 cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta cttgcattac agtttacgaa
     4741 ccgaacaggc ttatgtcaac tgggtttgtg ccttcatccg tttccacggt gtgcgtcacc
     4801 cggcaacctt gggcagcagc gaagtcgagg catttctgtc ctggctggcg aacgagcgca
     4861 aggtttcggt ctccacgcat cgtcaggcat tggcggcctt gctgttcttc tacggcaagg
     4921 tgctgtgcac ggatctgccc ttgcttcagg agatcggtag acctcggccg tcgcggcgct
     4981 tgccggtggt gctgaccccg gatgaagtgg ttcgcatcct cggttttctg gaaggcgagc
     5041 atcgtttgtt cgcccaggac tctagc
//
