LOCUS       sequence-243318-        5334 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculovirus vector for parallel SLIC cloning containing N-terminal 
            TwinStrep with PreScission Cleavage site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5334)
  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 5334)
  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: pCoofy51
FEATURES             Location/Qualifiers
     source          1..5334
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(517..522,523..539,540..545))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        897..988
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     913..930
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             join(1109..1114,1115..1138,1139..1174,1175..1198)
                     /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"
     CDS             1208..1231
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     RBS             1347..1355
                     /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             1357..1665
                     /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     1544..1563
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    1817..1951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1854..1873)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1908..1927
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1980..2145
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2329..2783
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2416..2435)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2625..2646
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2809..2913
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2914..2982,2983..3774)
                     /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(3132..3151)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3945..4533
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4434..4453
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4687..4704
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4838..5062
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(join(5129..5334,1..328))
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 catagatata gatctcacta cgcggctgct caaacttggg cagaacgtaa gccgcgagag
       61 cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat gaatgtctta ctacggagca
      121 agttcccgag gtaatcggag tccggctgat gttgggagta ggtggctacg tctccgaact
      181 cacgaccgaa aagatcaaga gcagcccgca tggatttgac ttggtcaggg ccgagcctac
      241 atgtgcgaat gatgcccata cttgagccac ctaactttgt tttagggcga ctgccctgct
      301 gcgtaacatc gttgctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc
      361 acggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacaaa aaaacagtca
      421 taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt caaggttctg
      481 gaccagttgc gtgagcgcat acgctacttg cattacagtt tacgaaccga acaggcttat
      541 gtcaactggg ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc aaccttgggc
      601 agcagcgaag tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt ttcggtctcc
      661 acgcatcgtc aggcattggc ggccttgctg ttcttctacg gcaaggtgct gtgcacggat
      721 ctgccctggc ttcaggagat cggaagacct cggccgtcgc ggcgcttgcc ggtggtgctg
      781 accccggatg aagtggttcg catcctcggt tttctggaag gcgagcatcg tttgttcgcc
      841 caggactcta gctatagttc tagtggttgg ctacgtatac tccggaatat taatagatca
      901 tggagataat taaaatgata accatctcgc aaataaataa gtattttact gttttcgtaa
      961 cagttttgta ataaaaaaac ctataaatat tccggattat tcataccgtc ccaccatcgg
     1021 gcgcggatcc cggtccgaag cgcgcggaat tcaaaggcct acgtcgacga gctcactagt
     1081 cgcggccgct ttcgaatcta gaatggggag cgcttggagc cacccgcagt tcgaaaaagg
     1141 tggaggttct ggcggtggat cgggaggttc agcgtggagc cacccgcagt tcgagaaaac
     1201 cgcgggtctg gaagttctgt tccaggggcc catggactaa tgaggtaccg gatccgaatt
     1261 cgagctccgt cgacaagctt gatatcgaat tcctgcagcc caaactgcag ttgacaacat
     1321 aaaaactttg tgttatactt gtaacgtaag gaggtaatga ttcagtttaa ggtttacacc
     1381 tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat tattgacacg
     1441 cccgggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga taaagtctcc
     1501 cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat gaccaccgat
     1561 atggccagtg tgccggtctc cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa
     1621 aatgacatca aaaacgccat taacctgatg ttctggggaa tataagcttc tcgaggcatg
     1681 cggtaccaag cttgtcgaga agtactagag gatcataatc agccatacca catttgtaga
     1741 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     1801 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     1861 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     1921 actcatcaat gtatcttatc atgtctggat ctgatcactg cttgagccta ggagatccga
     1981 accagataag tgaaatctag ttccaaacta ttttgtcatt tttaattttc gtattagctt
     2041 acgacgctac acccagttcc catctatttt gtcactcttc cctaaataat ccttaaaaac
     2101 tccatttcca cccctcccag ttcccaacta ttttgtccgc ccacagcggg gcatttttct
     2161 tcctgttatg tttttaatca aacatcctgc caactccatg tgacaaaccg tcatcttcgg
     2221 ctactttttc tctgtcacag aatgaaaatt tttctgtcat ctcttcgtta ttaatgtttg
     2281 taattgactg aatatcaacg cttatttgca gcctgaatgg cgaatgggac gcgccctgta
     2341 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2401 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2461 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2521 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     2581 agacggtttt tcgccctttg acgttggagt ccacgttctt aatagtggac tcttgttcca
     2641 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2701 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     2761 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     2821 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2881 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     2941 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     3001 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     3061 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     3121 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     3181 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     3241 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     3301 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     3361 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     3421 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     3481 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     3541 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3601 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     3661 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3721 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     3781 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3841 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3901 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3961 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4021 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4081 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4141 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4201 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4261 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4321 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4381 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4441 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4501 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4561 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4621 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4681 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     4741 acgcatctgt gcggtatttc acaccgcata gaccagccgc gtaacctggc aaaatcggtt
     4801 acggttgagt aataaatgga tgccctgcgt aagcgggtgt gggcggacaa taaagtctta
     4861 aactgaacaa aatagatcta aactatgaca ataaagtctt aaactagaca gaatagttgt
     4921 aaactgaaat cagtccagtt atgctgtgaa aaagcatact ggacttttgt tatggctaaa
     4981 gcaaactctt cattttctga agtgcaaatt gcccgtcgta ttaaagaggg gcgtggccaa
     5041 gggcatggta aagactatat tcgcggcgtt gtgacaattt accgaacaac tccgcggccg
     5101 ggaagccgat ctcggcttga acgaattgtt aggtggcggt acttgggtcg atatcaaagt
     5161 gcatcacttc ttcccgtatg cccaactttg tatagagagc cactgcggga tcgtcaccgt
     5221 aatctgcttg cacgtagatc acataagcac caagcgcgtt ggcctcatgc ttgaggagat
     5281 tgatgagcgc ggtggcaatg ccctgcctcc ggtgctcgcc ggagactgcg agat
//
