LOCUS       sequence-420505-        7422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of human Mcm2 N-terminally fused to mVenus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7422)
  TITLE     Pospiech Lab Plasmids 2023
REFERENCE   2  (bases 1 to 7422)
  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-Mcm2
FEATURES             Location/Qualifiers
     source          1..7422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        220..523
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        524..727
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     678..698
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      766..775
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(772..774,775..777,778..1488)
                     /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
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(817..838)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1078..1097)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1425..1446
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4369..4490
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4406..4425)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4460..4479
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4497..4952)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4634..4655)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4846..4865
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4979..5083
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5085..5442
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5085..5276
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5108..5128)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5293..5428
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5477..6271
                     /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             5477..6271
                     /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(5531..5550)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6141..6160
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6459..6478)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6503..6550
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(6879..7422,1..45)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7368..7387
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
       61 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
      121 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat
      181 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      241 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      301 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      361 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      421 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      481 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      541 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      601 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      661 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      721 cagagctggt ttagtgaacc gtcagatccg ctagcgctac cggtcgccac catggtgagc
      781 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
      841 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
      901 accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
      961 accctgggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     1021 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1081 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1141 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1201 tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
     1261 gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac
     1321 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1381 taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1441 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc cggactcaga
     1501 tctcgagctc aagcttcgaa ttcgatggcg gaatcatcgg aatccttcac catggcatcc
     1561 agcccggccc agcgtcggcg aggcaatgat cctctcacct ccagccctgg ccgaagctcc
     1621 cggcgtactg atgccctcac ctccagccct ggccgtgacc ttccaccatt tgaggatgag
     1681 tccgaggggc tcctaggcac agaggggccc ctggaggaag aagaggatgg agaggagctc
     1741 attggagatg gcatggaaag ggactaccgc gccatcccag agctggacgc ctatgaggcc
     1801 gagggactgg ctctggatga tgaggacgta gaggagctga cggccagtca gagggaggca
     1861 gcagagcggg ccatgcggca gcgtgaccgg gaggctggcc ggggcctggg ccgcatgcgc
     1921 cgtgggctcc tgtatgacag cgatgaggag gacgaggagc gccctgcccg caagcgccgc
     1981 caggtggagc gggccacgga ggacggcgag gaggacgagg agatgattga gagcatcgag
     2041 aacctggagg atctcaaagg ccactctgtg cgcgagtggg tgagcatggc gggcccccgg
     2101 ctggagatcc accaccgctt caagaacttc ctgcgcactc acgtcgacag ccacggccac
     2161 aacgtcttca aggagcgcat cagcgacatg tgcaaagaga accgtgagag cctggtggtg
     2221 aactatgagg acttggcagc cagggagcac gtgctggcct acttcctgcc tgaggcaccg
     2281 gcggagctgc tgcagatctt tgatgaggct gccctggagg tggtactggc catgtacccc
     2341 aagtacgacc gcatcaccaa ccacatccat gtccgcatct cccacctgcc tctggtggag
     2401 gagctgcgct cgctgaggca gctgcatctg aaccagctga tccgcaccag tggggtggtg
     2461 accagctgca ctggcgtcct gccccagctc agcatggtca agtacaactg caacaagtgc
     2521 aatttcgtcc tgggtccttt ctgccagtcc cagaaccagg aggtgaaacc aggctcctgt
     2581 cctgagtgcc agtcggccgg cccctttgag gtcaacatgg aggagaccat ctatcagaac
     2641 taccagcgta tccgaatcca ggagagtcca ggcaaagtgg cggctggccg gctgccccgc
     2701 tccaaggacg ccattctcct cgcagatctg gtggacagct gcaagccagg agacgagata
     2761 gagctgactg gcatctatca caacaactat gatggctccc tcaacactgc caatggcttc
     2821 cctgtctttg ccactgtcat cctagccaac cacgtggcca agaaggacaa caaggttgct
     2881 gtaggggaac tgaccgatga agatgtgaag atgatcacta gcctctccaa ggatcagcag
     2941 atcggagaga agatctttgc cagcattgct ccttccatct atggtcatga agacatcaag
     3001 agaggcctgg ctctggccct gttcggaggg gagcccaaaa acccaggtgg caagcacaag
     3061 gtacgtggtg atatcaacgt gctcttgtgc ggagaccctg gcacagcgaa gtcgcagttt
     3121 ctcaagtata ttgagaaagt gtccagccga gccatcttca ccactggcca gggggcgtcg
     3181 gctgtgggcc tcacggcgta tgtccagcgg caccctgtca gcagggagtg gaccttggag
     3241 gctggggccc tggttctggc tgaccgagga gtgtgtctca ttgatgaatt tgacaagatg
     3301 aatgaccagg acagaaccag catccatgag gccatggagc aacagagcat ctccatctcg
     3361 aaggctggca tcgtcacctc cctgcaggct cgctgcacgg tcattgctgc cgccaacccc
     3421 ataggagggc gctacgaccc ctcgctgact ttctctgaga acgtggacct cacagagccc
     3481 atcatctcac gctttgacat cctgtgtgtg gtgagggaca ccgtggaccc agtccaggac
     3541 gagatgctgg cccgcttcgt ggtgggcagc cacgtcagac accaccccag caacaaggag
     3601 gaggaggggc tggccaatgg cagcgctgct gagcccgcca tgcccaacac gtatggcgtg
     3661 gagcccctgc cccaggaggt cctgaagaag tacatcatct acgccaagga gagggtccac
     3721 ccgaagctca accagatgga ccaggacaag gtggccaaga tgtacagtga cctgaggaaa
     3781 gaatctatgg cgacaggcag catccccatt acggtgcggc acatcgagtc catgatccgc
     3841 atggcggagg cccacgcgcg catccatctg cgggactatg tgatcgaaga cgacgtcaac
     3901 atggccatcc gcgtgatgct ggagagcttc atagacacac agaagttcag cgtcatgcgc
     3961 agcatgcgca agacttttgc ccgctacctt tcattccggc gtgacaacaa tgagctgttg
     4021 ctcttcatac tgaagcagtt agtggcagag caggtgacat atcagcgcaa ccgctttggg
     4081 gcccagcagg acactattga ggtccctgag aaggacttgg tggataaggc tcgtcagatc
     4141 aacatccaca acctctctgc attttatgac agtgagctct tcaggatgaa caagttcagc
     4201 cacgacctga aaaggaaaat gatcctgcag cagttctgag gatccaccgg atctagataa
     4261 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     4321 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     4381 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4441 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat
     4501 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     4561 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     4621 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     4681 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     4741 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     4801 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     4861 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     4921 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     4981 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5041 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     5101 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     5161 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     5221 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     5281 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5341 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     5401 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     5461 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     5521 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     5581 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5641 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5701 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5761 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5821 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5881 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5941 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     6001 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     6061 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     6121 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     6181 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     6241 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     6301 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     6361 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     6421 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     6481 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     6541 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     6601 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     6661 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     6721 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6781 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6841 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6901 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6961 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7021 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7081 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7141 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7201 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7261 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7321 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7381 tatctttata gtcctgtcgg gtttcgccac ctctgacttg ag
//
