LOCUS       sequence-243586-        5563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculovirus vector for parallel SLIC cloning containing 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 5563)
  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 5563)
  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: pCoofy63
FEATURES             Location/Qualifiers
     source          1..5563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     16..33
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  167..391
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(458..991)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1161..1166,1167..1183,1184..1189))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        1541..1632
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1557..1574
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             1712..1729
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             2069..2077
                     /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             2079..2387
                     /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     2266..2285
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2391..2420
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             join(2424..2429,2430..2453,2454..2489,2490..2513)
                     /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    2709..2843
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2746..2765)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2800..2819
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2872..3037
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3221..3675
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3308..3327)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3517..3538
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3701..3805
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3806..3874,3875..4666)
                     /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(4024..4043)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4837..5425
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5326..5345
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
       61 tttctcctta cgcatctgtg cggtatttca caccgcatag accagccgcg taacctggca
      121 aaatcggtta cggttgagta ataaatggat gccctgcgta agcgggtgtg ggcggacaat
      181 aaagtcttaa actgaacaaa atagatctaa actatgacaa taaagtctta aactagacag
      241 aatagttgta aactgaaatc agtccagtta tgctgtgaaa aagcatactg gacttttgtt
      301 atggctaaag caaactcttc attttctgaa gtgcaaattg cccgtcgtat taaagagggg
      361 cgtggccaag ggcatggtaa agactatatt cgcggcgttg tgacaattta ccgaacaact
      421 ccgcggccgg gaagccgatc tcggcttgaa cgaattgtta ggtggcggta cttgggtcga
      481 tatcaaagtg catcacttct tcccgtatgc ccaactttgt atagagagcc actgcgggat
      541 cgtcaccgta atctgcttgc acgtagatca cataagcacc aagcgcgttg gcctcatgct
      601 tgaggagatt gatgagcgcg gtggcaatgc cctgcctccg gtgctcgccg gagactgcga
      661 gatcatagat atagatctca ctacgcggct gctcaaactt gggcagaacg taagccgcga
      721 gagcgccaac aaccgcttct tggtcgaagg cagcaagcgc gatgaatgtc ttactacgga
      781 gcaagttccc gaggtaatcg gagtccggct gatgttggga gtaggtggct acgtctccga
      841 actcacgacc gaaaagatca agagcagccc gcatggattt gacttggtca gggccgagcc
      901 tacatgtgcg aatgatgccc atacttgagc cacctaactt tgttttaggg cgactgccct
      961 gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt
     1021 gctgcttgga tgcccgaggc atagactgta caaaaaaaca gtcataacaa gccatgaaaa
     1081 ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc
     1141 gcatacgcta cttgcattac agtttacgaa ccgaacaggc ttatgtcaac tgggttcgtg
     1201 ccttcatccg tttccacggt gtgcgtcacc cggcaacctt gggcagcagc gaagtcgagg
     1261 catttctgtc ctggctggcg aacgagcgca aggtttcggt ctccacgcat cgtcaggcat
     1321 tggcggcctt gctgttcttc tacggcaagg tgctgtgcac ggatctgccc tggcttcagg
     1381 agatcggaag acctcggccg tcgcggcgct tgccggtggt gctgaccccg gatgaagtgg
     1441 ttcgcatcct cggttttctg gaaggcgagc atcgtttgtt cgcccaggac tctagctata
     1501 gttctagtgg ttggctacgt atactccgga atattaatag atcatggaga taattaaaat
     1561 gataaccatc tcgcaaataa ataagtattt tactgttttc gtaacagttt tgtaataaaa
     1621 aaacctataa atattccgga ttattcatac cgtcccacca tcgggcgcgg atcccggtcc
     1681 gaagcgcgcg gaattcatgg gccgcggaag ccaccatcac caccatcacg gctctggaat
     1741 gagcgactcc gaggtgaacc aggaggccaa gcccgaggtg aagcccgagg tgaagcccga
     1801 gacccacatc aacctgaagg tgtccgacgg ctcctccgag attttcttca agatcaagaa
     1861 gaccaccccc ctgcgccgcc tgatggaggc cttcgccaag cgccagggca aggagatgga
     1921 ctccctgacc ttcctgtacg acggcatcga gatccaggcc gaccagaccc ccgaggacct
     1981 ggacatggag gacaacgaca tcatcgaggc ccaccgcgag cagatcggcg gattgacaac
     2041 ataaaaactt tgtgttatac ttgtaacgta aggaggtaat gattcagttt aaggtttaca
     2101 cctataaaag agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca
     2161 cgcccgggcg acggatggtg atccccctgg ccagtgcacg tctgctgtca gataaagtct
     2221 cccgtgaact ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg
     2281 atatggccag tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg
     2341 aaaatgacat caaaaacgcc attaacctga tgttctgggg aatataagag catcatcatc
     2401 atcaccacca ccaccaccac tgaagcgctt ggagccaccc gcagttcgaa aaaggtggag
     2461 gttctggcgg tggatcggga ggttcagcgt ggagccaccc gcagttcgag aaataggaac
     2521 cggaagcgtg agcggccgct ttcgaatcta gagcctgcag tctcgaggca tgcggtacca
     2581 agcttgtcga gaagtactag aggatcataa tcagccatac cacatttgta gaggttttac
     2641 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     2701 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2761 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2821 atgtatctta tcatgtctgg atctgatcac tgcttgagcc taggagatcc gaaccagata
     2881 agtgaaatct agttccaaac tattttgtca tttttaattt tcgtattagc ttacgacgct
     2941 acacccagtt cccatctatt ttgtcactct tccctaaata atccttaaaa actccatttc
     3001 cacccctccc agttcccaac tattttgtcc gcccacagcg gggcattttt cttcctgtta
     3061 tgtttttaat caaacatcct gccaactcca tgtgacaaac cgtcatcttc ggctactttt
     3121 tctctgtcac agaatgaaaa tttttctgtc atctcttcgt tattaatgtt tgtaattgac
     3181 tgaatatcaa cgcttatttg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
     3241 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3301 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3361 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3421 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3481 tttcgccctt tgacgttgga gtccacgttc ttaatagtgg actcttgttc caaactggaa
     3541 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3601 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3661 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3721 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3781 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3841 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3901 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3961 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4021 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4081 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4141 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4201 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4261 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4321 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4381 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4441 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4501 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4561 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4621 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4681 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4741 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4801 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4861 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4921 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4981 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5041 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5101 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5161 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5221 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5281 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5341 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5401 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5461 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     5521 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agc
//
