LOCUS       sequence-243583-        5927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculovirus vector for parallel SLIC cloning containing N-terminal 
            His6-eGFP with PreScission Cleavage site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5927)
  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 5927)
  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: pCoofy60
FEATURES             Location/Qualifiers
     source          1..5927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  84..308
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(375..908)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1078..1083,1084..1100,1101..1106))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        1458..1549
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1474..1491
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             1610..1627
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(1628..1630,1631..2341)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLKE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1670..1691)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1931..1950)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2372..2395
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     RBS             2511..2519
                     /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             2521..2829
                     /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     2708..2727
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    2990..3124
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3027..3046)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3081..3100
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3153..3318
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3502..3956
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3589..3608)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3798..3819
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3982..4086
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4087..4155,4156..4947)
                     /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(4305..4324)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5118..5706
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5607..5626
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5860..5877
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tatttcacac cgcatagacc agccgcgtaa cctggcaaaa tcggttacgg ttgagtaata
       61 aatggatgcc ctgcgtaagc gggtgtgggc ggacaataaa gtcttaaact gaacaaaata
      121 gatctaaact atgacaataa agtcttaaac tagacagaat agttgtaaac tgaaatcagt
      181 ccagttatgc tgtgaaaaag catactggac ttttgttatg gctaaagcaa actcttcatt
      241 ttctgaagtg caaattgccc gtcgtattaa agaggggcgt ggccaagggc atggtaaaga
      301 ctatattcgc ggcgttgtga caatttaccg aacaactccg cggccgggaa gccgatctcg
      361 gcttgaacga attgttaggt ggcggtactt gggtcgatat caaagtgcat cacttcttcc
      421 cgtatgccca actttgtata gagagccact gcgggatcgt caccgtaatc tgcttgcacg
      481 tagatcacat aagcaccaag cgcgttggcc tcatgcttga ggagattgat gagcgcggtg
      541 gcaatgccct gcctccggtg ctcgccggag actgcgagat catagatata gatctcacta
      601 cgcggctgct caaacttggg cagaacgtaa gccgcgagag cgccaacaac cgcttcttgg
      661 tcgaaggcag caagcgcgat gaatgtctta ctacggagca agttcccgag gtaatcggag
      721 tccggctgat gttgggagta ggtggctacg tctccgaact cacgaccgaa aagatcaaga
      781 gcagcccgca tggatttgac ttggtcaggg ccgagcctac atgtgcgaat gatgcccata
      841 cttgagccac ctaactttgt tttagggcga ctgccctgct gcgtaacatc gttgctgctc
      901 cataacatca aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata
      961 gactgtacaa aaaaacagtc ataacaagcc atgaaaaccg ccactgcgcc gttaccaccg
     1021 ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca tacgctactt gcattacagt
     1081 ttacgaaccg aacaggctta tgtcaactgg gttcgtgcct tcatccgttt ccacggtgtg
     1141 cgtcacccgg caaccttggg cagcagcgaa gtcgaggcat ttctgtcctg gctggcgaac
     1201 gagcgcaagg tttcggtctc cacgcatcgt caggcattgg cggccttgct gttcttctac
     1261 ggcaaggtgc tgtgcacgga tctgccctgg cttcaggaga tcggaagacc tcggccgtcg
     1321 cggcgcttgc cggtggtgct gaccccggat gaagtggttc gcatcctcgg ttttctggaa
     1381 ggcgagcatc gtttgttcgc ccaggactct agctatagtt ctagtggttg gctacgtata
     1441 ctccggaata ttaatagatc atggagataa ttaaaatgat aaccatctcg caaataaata
     1501 agtattttac tgttttcgta acagttttgt aataaaaaaa cctataaata ttccggatta
     1561 ttcataccgt cccaccatcg ggcgcggatc ccggtccgaa accatgaaac atcaccatca
     1621 ccatcacgtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1681 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1741 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1801 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     1861 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1921 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1981 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2041 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     2101 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     2161 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     2221 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     2281 ggtccttaag gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     2341 ggggggtggt ggctccggtg gcggaggtac cctggaagtt ctgttccagg ggcccatgga
     2401 ctaatgaggt accggatccg aattcgagct ccgtcgacaa gcttgatatc gaattcctgc
     2461 agcccaaact gcagttgaca acataaaaac tttgtgttat acttgtaacg taaggaggta
     2521 atgattcagt ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat
     2581 gtacagagtg atattattga cacgcccggg cgacggatgg tgatccccct ggccagtgca
     2641 cgtctgctgt cagataaagt ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa
     2701 agctggcgca tgatgaccac cgatatggcc agtgtgccgg tctccgttat cggggaagaa
     2761 gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg
     2821 ggaatataag cttgcggccg cactcgaggc atgcggtacc aagcttgtcg agaagtacta
     2881 gaggatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     2941 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     3001 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3061 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3121 gatctgatca ctgcttgagc ctaggagatc cgaaccagat aagtgaaatc tagttccaaa
     3181 ctattttgtc atttttaatt ttcgtattag cttacgacgc tacacccagt tcccatctat
     3241 tttgtcactc ttccctaaat aatccttaaa aactccattt ccacccctcc cagttcccaa
     3301 ctattttgtc cgcccacagc ggggcatttt tcttcctgtt atgtttttaa tcaaacatcc
     3361 tgccaactcc atgtgacaaa ccgtcatctt cggctacttt ttctctgtca cagaatgaaa
     3421 atttttctgt catctcttcg ttattaatgt ttgtaattga ctgaatatca acgcttattt
     3481 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3541 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3601 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3661 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3721 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3781 agtccacgtt cttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3841 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3901 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
     3961 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4021 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4081 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4141 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4201 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4261 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4321 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4381 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4441 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     4501 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     4561 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     4621 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     4681 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     4741 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     4801 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     4861 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     4921 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     4981 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5041 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5101 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5161 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5221 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5281 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5341 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5401 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5461 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5521 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5581 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5641 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5701 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5761 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     5821 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     5881 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcgg
//
