LOCUS       sequence-243581-        5537 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculovirus vector for parallel SLIC cloning containing N-terminal 
            TrxA-His6 with PreScission Cleavage site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5537)
  AUTHORS   Scholz J, Besir H, Strasser C, Suppmann S
  TITLE     A new method to customize protein expression vectors for fast, 
            efficient and background free parallel cloning.
  JOURNAL   BMC Biotechnol. 2013 Feb 14;13(1):12.
  PUBMED    23410102
REFERENCE   2  (bases 1 to 5537)
  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: "BMC 
            Biotechnol."; date: "2013-02-14"; volume: "13(1)"; pages: "12"
COMMENT     Sequence Label: pCoofy59
FEATURES             Location/Qualifiers
     source          1..5537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      216..804
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     705..724
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     958..975
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  1109..1333
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1400..1933)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2103..2108,2109..2125,2126..2131))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        2483..2574
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2499..2516
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2629..2958
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MKSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILD
                     EIADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEF
                     LDANLA"
     CDS             2980..2997
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3007..3030
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     RBS             3146..3154
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             3156..3464
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MIQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSD
                     KVSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3343..3362
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    3625..3759
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3662..3681)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3716..3735
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3788..3953
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4137..4591
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4224..4243)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4433..4454
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4617..4721
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4722..4790,4791..5537,1..45)
                     /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(4940..4959)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
       61 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
      121 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
      181 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
      241 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
      301 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
      361 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
      421 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
      481 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
      541 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
      601 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
      661 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
      721 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
      781 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
      841 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
      901 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
      961 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     1021 tgcggtattt cacaccgcat agaccagccg cgtaacctgg caaaatcggt tacggttgag
     1081 taataaatgg atgccctgcg taagcgggtg tgggcggaca ataaagtctt aaactgaaca
     1141 aaatagatct aaactatgac aataaagtct taaactagac agaatagttg taaactgaaa
     1201 tcagtccagt tatgctgtga aaaagcatac tggacttttg ttatggctaa agcaaactct
     1261 tcattttctg aagtgcaaat tgcccgtcgt attaaagagg ggcgtggcca agggcatggt
     1321 aaagactata ttcgcggcgt tgtgacaatt taccgaacaa ctccgcggcc gggaagccga
     1381 tctcggcttg aacgaattgt taggtggcgg tacttgggtc gatatcaaag tgcatcactt
     1441 cttcccgtat gcccaacttt gtatagagag ccactgcggg atcgtcaccg taatctgctt
     1501 gcacgtagat cacataagca ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg
     1561 cggtggcaat gccctgcctc cggtgctcgc cggagactgc gagatcatag atatagatct
     1621 cactacgcgg ctgctcaaac ttgggcagaa cgtaagccgc gagagcgcca acaaccgctt
     1681 cttggtcgaa ggcagcaagc gcgatgaatg tcttactacg gagcaagttc ccgaggtaat
     1741 cggagtccgg ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat
     1801 caagagcagc ccgcatggat ttgacttggt cagggccgag cctacatgtg cgaatgatgc
     1861 ccatacttga gccacctaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc
     1921 tgctccataa catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag
     1981 gcatagactg tacaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac
     2041 caccgctgcg ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt
     2101 acagtttacg aaccgaacag gcttatgtca actgggttcg tgccttcatc cgtttccacg
     2161 gtgtgcgtca cccggcaacc ttgggcagca gcgaagtcga ggcatttctg tcctggctgg
     2221 cgaacgagcg caaggtttcg gtctccacgc atcgtcaggc attggcggcc ttgctgttct
     2281 tctacggcaa ggtgctgtgc acggatctgc cctggcttca ggagatcgga agacctcggc
     2341 cgtcgcggcg cttgccggtg gtgctgaccc cggatgaagt ggttcgcatc ctcggttttc
     2401 tggaaggcga gcatcgtttg ttcgcccagg actctagcta tagttctagt ggttggctac
     2461 gtatactccg gaatattaat agatcatgga gataattaaa atgataacca tctcgcaaat
     2521 aaataagtat tttactgttt tcgtaacagt tttgtaataa aaaaacctat aaatattccg
     2581 gattattcat accgtcccac catcgggcgc ggatcccggt ccgaaaccat gaaaagcgat
     2641 aaaattattc acctgactga cgacagtttt gacacggatg tactcaaagc ggacggggcg
     2701 atcctcgtcg atttctgggc agagtggtgc ggtccgtgca aaatgatcgc cccgattctg
     2761 gatgaaatcg ctgacgaata tcagggcaaa ctgaccgttg caaaactgaa catcgatcaa
     2821 aaccctggca ctgcgccgaa atatggcatc cgtggtatcc cgactctgct gctgttcaaa
     2881 aacggtgaag tggcggcaac caaagtgggt gcactgtcta aaggtcagtt gaaagagttc
     2941 ctcgacgcta acctggccgg ttctggttct ggccatatgc accatcatca tcatcattct
     3001 tctggtctgg aagttctgtt ccaggggccc atggactaat gaggtaccgg atccgaattc
     3061 gagctccgtc gacaagcttg atatcgaatt cctgcagccc aaactgcagt tgacaacata
     3121 aaaactttgt gttatacttg taacgtaagg aggtaatgat tcagtttaag gtttacacct
     3181 ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc
     3241 ccgggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtctccc
     3301 gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata
     3361 tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa
     3421 atgacatcaa aaacgccatt aacctgatgt tctggggaat ataagcttgc ggccgcactc
     3481 gaggcatgcg gtaccaagct tgtcgagaag tactagagga tcataatcag ccataccaca
     3541 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     3601 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     3661 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3721 ttgtccaaac tcatcaatgt atcttatcat gtctggatct gatcactgct tgagcctagg
     3781 agatccgaac cagataagtg aaatctagtt ccaaactatt ttgtcatttt taattttcgt
     3841 attagcttac gacgctacac ccagttccca tctattttgt cactcttccc taaataatcc
     3901 ttaaaaactc catttccacc cctcccagtt cccaactatt ttgtccgccc acagcggggc
     3961 atttttcttc ctgttatgtt tttaatcaaa catcctgcca actccatgtg acaaaccgtc
     4021 atcttcggct actttttctc tgtcacagaa tgaaaatttt tctgtcatct cttcgttatt
     4081 aatgtttgta attgactgaa tatcaacgct tatttgcagc ctgaatggcg aatgggacgc
     4141 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4201 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4261 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4321 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     4381 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttaa tagtggactc
     4441 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4501 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4561 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     4621 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4681 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4741 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4801 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4861 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4921 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4981 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5041 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5101 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5161 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5221 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5281 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5341 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5401 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5461 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5521 ctatggatga acgaaat
//
