LOCUS       sequence-243584-        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"
     primer_bind     69..86
                     /label=L4440
                     /note="L4440 vector, forward primer"
     enhancer        133..512
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        513..716
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     667..687
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     713..737
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1218..1237
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1389..1406
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1746..1754
                     /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             1756..2064
                     /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     1943..1962
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2068..2097
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             join(2101..2106,2107..2130,2131..2166,2167..2190)
                     /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    2260..2315
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2314..2333)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3398..3416)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3484..3588
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3589..3657,3658..4449)
                     /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(3807..3826)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4620..5208
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5109..5128
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
       61 ccgagcgcag cgagtcagtg agcgaggaag cgtacattta tattggctca tgtccaatat
      121 gaccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
      181 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      241 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      301 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      361 tggcagtaca tcaagtgtat catatgccaa gtccgccccc tattgacgtc aatgacggta
      421 aatggcccgc ctggcattat gcccagtaca tgaccttacg ggactttcct acttggcagt
      481 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacaccaatg
      541 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      601 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaat aaccccgccc
      661 cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt
      721 tagtgaaccg tcagatcctc actctcttcc gcatcgctgt ctgcgagggc cagctgttgg
      781 gctcgcggtt gaggacaaac tcttcgcggt ctttccagta ctcttggatc ggaaacccgt
      841 cggcctccga acggtactcc gccaccgagg gacctgagcg agtccgcatc gaccggatcg
      901 gaaaacctct cgagaaaggc gtctaaccag tcacagtcgc aaggtaggct gagcaccgtg
      961 gcgggcggca gcgggtggcg gtcggggttg tttctggcgg aggtgctgct gatgatgtaa
     1021 ttaaagtagg cggtcttgag acggcggatg gtcgaggtga ggtgtggcag gcttgagatc
     1081 cagctgttgg ggtgagtact ccctctcaaa agcgggcatt acttctgcgc taagattgtc
     1141 agtttccaaa aacgaggagg atttgatatt cacctggccc gatctggcca tacacttgag
     1201 tgacaatgac atccactttg cctttctctc cacaggtgtc cactcccagg tccaagttta
     1261 aacggatctc tagcgaattc atgaactttc tgctgtcctg ggtgcactgg tccctggccc
     1321 tgctgctgta cctgcaccac gccaagtggt cccaggccgc tcctatggcc gagggcggtg
     1381 gccagaacca ccatcaccac catcacggct ctggaatgag cgactccgag gtgaaccagg
     1441 aggccaagcc cgaggtgaag cccgaggtga agcccgagac ccacatcaac ctgaaggtgt
     1501 ccgacggctc ctccgagatt ttcttcaaga tcaagaagac cacccccctg cgccgcctga
     1561 tggaggcctt cgccaagcgc cagggcaagg agatggactc cctgaccttc ctgtacgacg
     1621 gcatcgagat ccaggccgac cagacccccg aggacctgga catggaggac aacgacatca
     1681 tcgaggccca ccgcgagcag atcggcggat tgacaacata aaaactttgt gttatacttg
     1741 taacgtaagg aggtaatgat tcagtttaag gtttacacct ataaaagaga gagccgttat
     1801 cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg gatggtgatc
     1861 cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg
     1921 catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc
     1981 gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt
     2041 aacctgatgt tctggggaat ataagagcat catcatcatc accaccacca ccaccactga
     2101 agcgcttgga gccacccgca gttcgaaaaa ggtggaggtt ctggcggtgg atcgggaggt
     2161 tcagcgtgga gccacccgca gttcgagaaa taggaaccgg aagcgtgagc ggccgctcga
     2221 ggccggcaag gccggatccc ccgacctcga cctctggcta ataaaggaaa tttattttca
     2281 ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa
     2341 atcatttggt cgagatccct cggagatctc tagctagagg atcgatcccc gccccggacg
     2401 aactaaacct gactacgaca tctctgcccc ttcttcgcgg ggcagtgcat gtaatccctt
     2461 cagttggttg gtacaacttg ccaactgaac cctaaacggg tagcatatgc ttcccgggta
     2521 gtagtatata ctatccagac taaccctaat tcaatagcat atgttaccca acgggaagca
     2581 tatgctatcg aattagggtt agtaaaaggg tcctaaggaa cagcgatgta ggtgggcggg
     2641 ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat tacacacact tgcgcctgag
     2701 cgccaagcac agggttgttg gtcctcatat tcacgaggtc gctgagagca cggtgggcta
     2761 atgttgccat gggtagcata tactacccaa atatctggat agcatatgct atcctaatct
     2821 atatctgggt agcataggct atcctaatct atatctgggt agcatatgct atcctaatct
     2881 atatctgggt agtatatgct atcctaattt atatctgggt agcataggct atcctaatct
     2941 atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct
     3001 gtatccgggt agcatatgct atcctaatag agattagggt agtatatgct atcctaattt
     3061 atatctgggt agcatatact acccaaatat ctggatagca tatgctatcc taatctatat
     3121 ctgggtagca tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat
     3181 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat
     3241 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     3301 ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc tcatgataag
     3361 ctgtcaaaca tgagaattaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     3421 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     3481 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3541 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     3601 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3661 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     3721 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     3781 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     3841 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     3901 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3961 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4021 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4081 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     4141 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4201 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4261 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     4321 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4381 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4441 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     4501 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4561 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4621 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4681 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4741 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4801 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4861 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4921 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4981 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5041 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5101 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5161 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5221 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5281 ttatcccctg att
//
