LOCUS       sequence-243578-        5200 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian 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 5200)
  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 5200)
  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: pCoofy65
FEATURES             Location/Qualifiers
     source          1..5200
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        42..421
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        422..625
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     576..596
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     622..646
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1127..1146
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1205..1222
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1562..1570
                     /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             1572..1880
                     /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     1759..1778
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1884..1913
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             join(1917..1922,1923..1946,1947..1982,1983..2006)
                     /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    2076..2131
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2130..2149)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3214..3232)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3300..3404
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3405..3473,3474..4265)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3623..3642)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4436..5024
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4925..4944
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5178..5195
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtacatttat attggctcat gtccaatatg accgccatgt tgacattgat tattgactag
       61 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      121 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      181 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      241 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      301 tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      361 gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      421 ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt
      481 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      541 ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg
      601 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcctca ctctcttccg
      661 catcgctgtc tgcgagggcc agctgttggg ctcgcggttg aggacaaact cttcgcggtc
      721 tttccagtac tcttggatcg gaaacccgtc ggcctccgaa cggtactccg ccaccgaggg
      781 acctgagcga gtccgcatcg accggatcgg aaaacctctc gagaaaggcg tctaaccagt
      841 cacagtcgca aggtaggctg agcaccgtgg cgggcggcag cgggtggcgg tcggggttgt
      901 ttctggcgga ggtgctgctg atgatgtaat taaagtaggc ggtcttgaga cggcggatgg
      961 tcgaggtgag gtgtggcagg cttgagatcc agctgttggg gtgagtactc cctctcaaaa
     1021 gcgggcatta cttctgcgct aagattgtca gtttccaaaa acgaggagga tttgatattc
     1081 acctggcccg atctggccat acacttgagt gacaatgaca tccactttgc ctttctctcc
     1141 acaggtgtcc actcccaggt ccaagtttaa acggatctct agcgaattca tgggccgcgg
     1201 aagccaccat caccaccatc acggctctgg aatgagcgac tccgaggtga accaggaggc
     1261 caagcccgag gtgaagcccg aggtgaagcc cgagacccac atcaacctga aggtgtccga
     1321 cggctcctcc gagattttct tcaagatcaa gaagaccacc cccctgcgcc gcctgatgga
     1381 ggccttcgcc aagcgccagg gcaaggagat ggactccctg accttcctgt acgacggcat
     1441 cgagatccag gccgaccaga cccccgagga cctggacatg gaggacaacg acatcatcga
     1501 ggcccaccgc gagcagatcg gcggattgac aacataaaaa ctttgtgtta tacttgtaac
     1561 gtaaggaggt aatgattcag tttaaggttt acacctataa aagagagagc cgttatcgtc
     1621 tgtttgtgga tgtacagagt gatattattg acacgcccgg gcgacggatg gtgatccccc
     1681 tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata
     1741 tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta
     1801 tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc
     1861 tgatgttctg gggaatataa gagcatcatc atcatcacca ccaccaccac cactgaagcg
     1921 cttggagcca cccgcagttc gaaaaaggtg gaggttctgg cggtggatcg ggaggttcag
     1981 cgtggagcca cccgcagttc gagaaatagg aaccggaagc gtgagcggcc gctcgaggcc
     2041 ggcaaggccg gatcccccga cctcgacctc tggctaataa aggaaattta ttttcattgc
     2101 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     2161 tttggtcgag atccctcgga gatctctagc tagaggatcg atccccgccc cggacgaact
     2221 aaacctgact acgacatctc tgccccttct tcgcggggca gtgcatgtaa tcccttcagt
     2281 tggttggtac aacttgccaa ctgaacccta aacgggtagc atatgcttcc cgggtagtag
     2341 tatatactat ccagactaac cctaattcaa tagcatatgt tacccaacgg gaagcatatg
     2401 ctatcgaatt agggttagta aaagggtcct aaggaacagc gatgtaggtg ggcgggccaa
     2461 gataggggcg cgattgctgc gatctggagg acaaattaca cacacttgcg cctgagcgcc
     2521 aagcacaggg ttgttggtcc tcatattcac gaggtcgctg agagcacggt gggctaatgt
     2581 tgccatgggt agcatatact acccaaatat ctggatagca tatgctatcc taatctatat
     2641 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     2701 ctgggtagta tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat
     2761 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat
     2821 ccgggtagca tatgctatcc taatagagat tagggtagta tatgctatcc taatttatat
     2881 ctgggtagca tatactaccc aaatatctgg atagcatatg ctatcctaat ctatatctgg
     2941 gtagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat ctatatctgg
     3001 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ttatatctgg
     3061 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     3121 gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctcat gataagctgt
     3181 caaacatgag aattaattct tgaagacgaa agggcctcgt gatacgccta tttttatagg
     3241 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     3301 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3361 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     3421 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3481 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     3541 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     3601 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
     3661 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     3721 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3781 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     3841 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3901 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     3961 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     4021 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     4081 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     4141 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     4201 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4261 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     4321 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4381 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4441 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4501 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4561 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4621 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4681 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4741 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4801 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4861 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4921 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4981 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5041 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5101 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     5161 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
//
