LOCUS       sequence-232398-        5293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian vector for parallel SLIC cloning containing the VEGF 
            signal sequence for secretion, 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 5293)
  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 5293)
  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: pCoofy62
FEATURES             Location/Qualifiers
     source          1..5293
                     /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             1298..1315
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1655..1663
                     /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             1665..1973
                     /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     1852..1871
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1977..2006
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             join(2010..2015,2016..2039,2040..2075,2076..2099)
                     /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    2169..2224
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2223..2242)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3307..3325)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3393..3497
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3498..3566,3567..4358)
                     /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(3716..3735)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4529..5117
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5018..5037
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5271..5288
                     /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 tgaactttct
     1201 gctgtcctgg gtgcactggt ccctggccct gctgctgtac ctgcaccacg ccaagtggtc
     1261 ccaggccgct cctatggccg agggcggtgg ccagaaccac catcaccacc atcacggctc
     1321 tggaatgagc gactccgagg tgaaccagga ggccaagccc gaggtgaagc ccgaggtgaa
     1381 gcccgagacc cacatcaacc tgaaggtgtc cgacggctcc tccgagattt tcttcaagat
     1441 caagaagacc acccccctgc gccgcctgat ggaggccttc gccaagcgcc agggcaagga
     1501 gatggactcc ctgaccttcc tgtacgacgg catcgagatc caggccgacc agacccccga
     1561 ggacctggac atggaggaca acgacatcat cgaggcccac cgcgagcaga tcggcggatt
     1621 gacaacataa aaactttgtg ttatacttgt aacgtaagga ggtaatgatt cagtttaagg
     1681 tttacaccta taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta
     1741 ttgacacgcc cgggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata
     1801 aagtctcccg tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga
     1861 ccaccgatat ggccagtgtg ccggtctccg ttatcgggga agaagtggct gatctcagcc
     1921 accgcgaaaa tgacatcaaa aacgccatta acctgatgtt ctggggaata taagagcatc
     1981 atcatcatca ccaccaccac caccactgaa gcgcttggag ccacccgcag ttcgaaaaag
     2041 gtggaggttc tggcggtgga tcgggaggtt cagcgtggag ccacccgcag ttcgagaaat
     2101 aggaaccgga agcgtgagcg gccgctcgag gccggcaagg ccggatcccc cgacctcgac
     2161 ctctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     2221 ctcactcgga aggacatatg ggagggcaaa tcatttggtc gagatccctc ggagatctct
     2281 agctagagga tcgatccccg ccccggacga actaaacctg actacgacat ctctgcccct
     2341 tcttcgcggg gcagtgcatg taatcccttc agttggttgg tacaacttgc caactgaacc
     2401 ctaaacgggt agcatatgct tcccgggtag tagtatatac tatccagact aaccctaatt
     2461 caatagcata tgttacccaa cgggaagcat atgctatcga attagggtta gtaaaagggt
     2521 cctaaggaac agcgatgtag gtgggcgggc caagataggg gcgcgattgc tgcgatctgg
     2581 aggacaaatt acacacactt gcgcctgagc gccaagcaca gggttgttgg tcctcatatt
     2641 cacgaggtcg ctgagagcac ggtgggctaa tgttgccatg ggtagcatat actacccaaa
     2701 tatctggata gcatatgcta tcctaatcta tatctgggta gcataggcta tcctaatcta
     2761 tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaattta
     2821 tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     2881 tatctgggta gtatatgcta tcctaatctg tatccgggta gcatatgcta tcctaataga
     2941 gattagggta gtatatgcta tcctaattta tatctgggta gcatatacta cccaaatatc
     3001 tggatagcat atgctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     3061 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     3121 tgggtagtat atgctatcct aatttatatc tgggtagcat aggctatcct aatctatatc
     3181 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatctgtatc
     3241 cgggtagcat atgctatcct catgataagc tgtcaaacat gagaattaat tcttgaagac
     3301 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     3361 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3421 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3481 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3541 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3601 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3661 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3721 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     3781 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3841 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3901 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3961 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4021 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4081 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4141 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4201 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4261 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4321 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4381 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4441 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4501 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4561 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4621 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4681 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4741 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4801 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4861 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4921 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4981 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5041 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5101 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5161 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5221 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     5281 tgagcgagga agc
//
