LOCUS       sequence-243594-        5693 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculovirus vector for parallel SLIC cloning containing gp67 signal 
            sequence, N-terminal His6-Smt3Star (SumoStar*) and different C-tag 
            options.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5693)
  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 5693)
  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: pCoofy64
FEATURES             Location/Qualifiers
     source          1..5693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        87..178
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     103..120
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     sig_peptide     297..356
                     /codon_start=1
                     /gene="Autographa californica gp64"
                     /product="signal sequence from the baculovirus envelope 
                     glycoprotein GP64"
                     /label=GP64 signal sequence
                     /translation="MVSAIVLYVLLAAAAHSAFA"
     CDS             369..386
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             726..734
                     /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             736..1044
                     /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     923..942
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1048..1077
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             join(1081..1086,1087..1110,1111..1146,1147..1170)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     polyA_signal    1366..1500
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1403..1422)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1457..1476
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1529..1694
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      1878..2332
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1965..1984)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2174..2195
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2358..2462
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2463..2531,2532..3323)
                     /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(2681..2700)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3494..4082
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3983..4002
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4236..4253
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4387..4611
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4678..5211)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5400..5405,5406..5422,5423..5428))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 tttctggaag gcgagcatcg tttgttcgcc caggactcta gctatagttc tagtggttgg
       61 ctacgtatac tccggaatat taatagatca tggagataat taaaatgata accatctcgc
      121 aaataaataa gtattttact gttttcgtaa cagttttgta ataaaaaaac ctataaatat
      181 tccggattat tcataccgtc ccaccatcgg gcgcggatcc cggtccgaag cgcgcggaat
      241 tcatgctact agtaaatcag tcacaccaag gcttcaataa ggaacacaca agcaagatgg
      301 taagcgctat tgttttatat gtgcttttgg cggcggcggc gcattctgcc tttgcggcag
      361 gtatgggtca ccatcaccac catcacggct ctggaatgag cgactccgag gtgaaccagg
      421 aggccaagcc cgaggtgaag cccgaggtga agcccgagac ccacatcaac ctgaaggtgt
      481 ccgacggctc ctccgagatt ttcttcaaga tcaagaagac cacccccctg cgccgcctga
      541 tggaggcctt cgccaagcgc cagggcaagg agatggactc cctgaccttc ctgtacgacg
      601 gcatcgagat ccaggccgac cagacccccg aggacctgga catggaggac aacgacatca
      661 tcgaggccca ccgcgagcag atcggcggat tgacaacata aaaactttgt gttatacttg
      721 taacgtaagg aggtaatgat tcagtttaag gtttacacct ataaaagaga gagccgttat
      781 cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg gatggtgatc
      841 cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg
      901 catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc
      961 gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt
     1021 aacctgatgt tctggggaat ataagagcat catcatcatc accaccacca ccaccactga
     1081 agcgcttgga gccacccgca gttcgaaaaa ggtggaggtt ctggcggtgg atcgggaggt
     1141 tcagcgtgga gccacccgca gttcgagaaa taggaaccgg aagcgtgagc ggccgctttc
     1201 gaatctagag cctgcagtct cgaggcatgc ggtaccaagc ttgtcgagaa gtactagagg
     1261 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     1321 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     1381 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     1441 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     1501 tgatcactgc ttgagcctag gagatccgaa ccagataagt gaaatctagt tccaaactat
     1561 tttgtcattt ttaattttcg tattagctta cgacgctaca cccagttccc atctattttg
     1621 tcactcttcc ctaaataatc cttaaaaact ccatttccac ccctcccagt tcccaactat
     1681 tttgtccgcc cacagcgggg catttttctt cctgttatgt ttttaatcaa acatcctgcc
     1741 aactccatgt gacaaaccgt catcttcggc tactttttct ctgtcacaga atgaaaattt
     1801 ttctgtcatc tcttcgttat taatgtttgt aattgactga atatcaacgc ttatttgcag
     1861 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     1921 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     1981 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2041 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2101 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2161 cacgttctta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     2221 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     2281 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     2341 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2401 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2461 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2521 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2581 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2641 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2701 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2761 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2821 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     2881 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     2941 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3001 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3061 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     3121 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3181 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3241 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3301 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     3361 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3421 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3481 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3541 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3601 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     3661 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3721 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3781 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3841 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3901 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3961 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4021 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4081 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4141 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     4201 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     4261 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag
     4321 accagccgcg taacctggca aaatcggtta cggttgagta ataaatggat gccctgcgta
     4381 agcgggtgtg ggcggacaat aaagtcttaa actgaacaaa atagatctaa actatgacaa
     4441 taaagtctta aactagacag aatagttgta aactgaaatc agtccagtta tgctgtgaaa
     4501 aagcatactg gacttttgtt atggctaaag caaactcttc attttctgaa gtgcaaattg
     4561 cccgtcgtat taaagagggg cgtggccaag ggcatggtaa agactatatt cgcggcgttg
     4621 tgacaattta ccgaacaact ccgcggccgg gaagccgatc tcggcttgaa cgaattgtta
     4681 ggtggcggta cttgggtcga tatcaaagtg catcacttct tcccgtatgc ccaactttgt
     4741 atagagagcc actgcgggat cgtcaccgta atctgcttgc acgtagatca cataagcacc
     4801 aagcgcgttg gcctcatgct tgaggagatt gatgagcgcg gtggcaatgc cctgcctccg
     4861 gtgctcgccg gagactgcga gatcatagat atagatctca ctacgcggct gctcaaactt
     4921 gggcagaacg taagccgcga gagcgccaac aaccgcttct tggtcgaagg cagcaagcgc
     4981 gatgaatgtc ttactacgga gcaagttccc gaggtaatcg gagtccggct gatgttggga
     5041 gtaggtggct acgtctccga actcacgacc gaaaagatca agagcagccc gcatggattt
     5101 gacttggtca gggccgagcc tacatgtgcg aatgatgccc atacttgagc cacctaactt
     5161 tgttttaggg cgactgccct gctgcgtaac atcgttgctg ctgcgtaaca tcgttgctgc
     5221 tccataacat caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca
     5281 tagactgtac aaaaaaacag tcataacaag ccatgaaaac cgccactgcg ccgttaccac
     5341 cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg catacgctac ttgcattaca
     5401 gtttacgaac cgaacaggct tatgtcaact gggttcgtgc cttcatccgt ttccacggtg
     5461 tgcgtcaccc ggcaaccttg ggcagcagcg aagtcgaggc atttctgtcc tggctggcga
     5521 acgagcgcaa ggtttcggtc tccacgcatc gtcaggcatt ggcggccttg ctgttcttct
     5581 acggcaaggt gctgtgcacg gatctgccct ggcttcagga gatcggaaga cctcggccgt
     5641 cgcggcgctt gccggtggtg ctgaccccgg atgaagtggt tcgcatcctc ggt
//
