LOCUS       sequence-243400-        5274 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian vector for parallel SLIC cloning containing N-terminal 
            His10-SumoStar (Life Sensors) with additional PreScission Cleavage 
            site and different C-tag options.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5274)
  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 5274)
  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: pCoofy55
FEATURES             Location/Qualifiers
     source          1..5274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     52..69
                     /label=L4440
                     /note="L4440 vector, forward primer"
     enhancer        116..495
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        496..699
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     650..670
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     696..720
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1201..1220
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1250..1279
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1289..1576
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated and modified)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /note="modified to prevent cleavage by eukaryotic SUMO 
                     proteases"
                     /translation="DSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRR
                     LMEAFAKRQGKEMDSLTFLYDGIEIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             1583..1606
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     RBS             1643..1651
                     /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             1653..1961
                     /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     1840..1859
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1965..1994
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             join(1998..2003,2004..2027,2028..2063,2064..2087)
                     /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             2091..2135
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    2224..2279
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2278..2297)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3362..3380)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3448..3552
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3553..3621,3622..4413)
                     /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(3771..3790)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4584..5172
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5073..5092
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
       61 gtgagcgagg aagcgtacat ttatattggc tcatgtccaa tatgaccgcc atgttgacat
      121 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      181 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      241 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      301 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      361 tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      421 tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc
      481 atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg atagcggttt
      541 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      601 caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc
      661 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc
      721 ctcactctct tccgcatcgc tgtctgcgag ggccagctgt tgggctcgcg gttgaggaca
      781 aactcttcgc ggtctttcca gtactcttgg atcggaaacc cgtcggcctc cgaacggtac
      841 tccgccaccg agggacctga gcgagtccgc atcgaccgga tcggaaaacc tctcgagaaa
      901 ggcgtctaac cagtcacagt cgcaaggtag gctgagcacc gtggcgggcg gcagcgggtg
      961 gcggtcgggg ttgtttctgg cggaggtgct gctgatgatg taattaaagt aggcggtctt
     1021 gagacggcgg atggtcgagg tgaggtgtgg caggcttgag atccagctgt tggggtgagt
     1081 actccctctc aaaagcgggc attacttctg cgctaagatt gtcagtttcc aaaaacgagg
     1141 aggatttgat attcacctgg cccgatctgg ccatacactt gagtgacaat gacatccact
     1201 ttgcctttct ctccacaggt gtccactccc aggtccaagt ttaaacatgc accatcacca
     1261 ccatcaccat catcatcact ccctgcagga ctcagaagtc aatcaagaag ctaagccaga
     1321 ggtcaagcca gaagtcaagc ctgagactca catcaattta aaggtgtccg atggatcttc
     1381 agagatcttc ttcaagatca aaaagaccac tcctttaaga aggctgatgg aagcgttcgc
     1441 taaaagacag ggtaaggaaa tggactcctt aacgttcttg tacgacggta ttgaaattca
     1501 agctgatcag acccctgaag atttggacat ggaggataac gatattattg aggctcatcg
     1561 cgaacagatt ggaggagaat tcctggaagt tctgttccag gggcccttga caacataaaa
     1621 actttgtgtt atacttgtaa cgtaaggagg taatgattca gtttaaggtt tacacctata
     1681 aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt gacacgcccg
     1741 ggcgacggat ggtgatcccc ctggccagtg cacgtctgct gtcagataaa gtctcccgtg
     1801 aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc accgatatgg
     1861 ccagtgtgcc ggtctccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg
     1921 acatcaaaaa cgccattaac ctgatgttct ggggaatata agagcatcat catcatcacc
     1981 accaccacca ccactgaagc gcttggagcc acccgcagtt cgaaaaaggt ggaggttctg
     2041 gcggtggatc gggaggttca gcgtggagcc acccgcagtt cgagaaatag aaagaaaccg
     2101 ctgctgctaa attcgaacgc cagcacatgg acagctagtc tagagggccc gtttctgcta
     2161 gcaagcttgc tagcggccgc tcgaggccgg caaggccgga tcccccgacc tcgacctctg
     2221 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     2281 tcggaaggac atatgggagg gcaaatcatt tggtcgagat ccctcggaga tctctagcta
     2341 gaggatcgat ccccgccccg gacgaactaa acctgactac gacatctctg ccccttcttc
     2401 gcggggcagt gcatgtaatc ccttcagttg gttggtacaa cttgccaact gaaccctaaa
     2461 cgggtagcat atgcttcccg ggtagtagta tatactatcc agactaaccc taattcaata
     2521 gcatatgtta cccaacggga agcatatgct atcgaattag ggttagtaaa agggtcctaa
     2581 ggaacagcga tgtaggtggg cgggccaaga taggggcgcg attgctgcga tctggaggac
     2641 aaattacaca cacttgcgcc tgagcgccaa gcacagggtt gttggtcctc atattcacga
     2701 ggtcgctgag agcacggtgg gctaatgttg ccatgggtag catatactac ccaaatatct
     2761 ggatagcata tgctatccta atctatatct gggtagcata ggctatccta atctatatct
     2821 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atttatatct
     2881 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
     2941 gggtagtata tgctatccta atctgtatcc gggtagcata tgctatccta atagagatta
     3001 gggtagtata tgctatccta atttatatct gggtagcata tactacccaa atatctggat
     3061 agcatatgct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     3121 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     3181 agtatatgct atcctaattt atatctgggt agcataggct atcctaatct atatctgggt
     3241 agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt
     3301 agcatatgct atcctcatga taagctgtca aacatgagaa ttaattcttg aagacgaaag
     3361 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     3421 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3481 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3541 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3601 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3661 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3721 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3781 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3841 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3901 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3961 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4021 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4081 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4141 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4201 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4261 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4321 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4381 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4441 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4501 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4561 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4621 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4681 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4741 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4801 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4861 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4921 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4981 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5041 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5101 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5161 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5221 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tatt
//
