LOCUS       sequence-420410-        5296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector backbone for coupled mammalian expression of N-terminal Venus
            fusion protein and another protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5296)
  TITLE     Pospiech Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5296)
  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"
COMMENT     Sequence Label: pmVen-IRES
FEATURES             Location/Qualifiers
     source          1..5296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        83..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      629..638
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(635..637,638..640,641..1348)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(680..701)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(941..960)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1288..1309
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1522..1539)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1749..1768
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    1926..1982
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2106..2227
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2143..2162)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2197..2216
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2234..2689)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2371..2392)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2583..2602
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2716..2820
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2822..3179
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2822..3013
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2845..2865)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3030..3165
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3214..4008
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3214..4008
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3268..3287)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3878..3897
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4196..4215)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4240..4287
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4616..5204
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5105..5124
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
      601 accgtcagat ccgctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt
      661 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
      721 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg
      781 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcct
      841 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
      901 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
      961 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1021 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1081 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg
     1141 ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat
     1201 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaagctgag
     1261 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1321 gatcactctc ggcatggacg agctgtacat tccgcccctc tccctccccc ccccctaacg
     1381 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     1441 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     1501 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     1561 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     1621 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     1681 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     1741 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     1801 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     1861 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     1921 gataaagatc tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat
     1981 ccaccggatc tagataactg atcataatca gccataccac atttgtagag gttttacttg
     2041 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     2101 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     2161 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     2221 tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     2281 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     2341 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     2401 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2461 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     2521 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     2581 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     2641 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     2701 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2761 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     2821 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     2881 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     2941 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3001 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3061 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     3121 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     3181 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3241 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3301 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3361 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     3421 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3481 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3541 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3601 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     3661 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     3721 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     3781 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     3841 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     3901 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     3961 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4021 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4081 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4141 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     4201 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     4261 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     4321 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     4381 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     4441 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     4501 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4561 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4621 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4681 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4741 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4801 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4861 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4921 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4981 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5041 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5101 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5161 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5221 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5281 cccctgattc tgtgga
//
