LOCUS       sequence-420427-        7823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of human Cdc45 C-terminally fused to mVenus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7823)
  TITLE     Pospiech Lab Plasmids 2023
REFERENCE   2  (bases 1 to 7823)
  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: pcDNA3-Cdc45-mVen
FEATURES             Location/Qualifiers
     source          1..7823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(60..79)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        251..630
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        631..834
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     785..805
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     879..898
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        879..897
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2657..2666
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2663..2665,2666..2668,2669..3382)
                     /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(2708..2729)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2969..2988)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3316..3337
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(3392..3410)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3393..3410)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3430..3447)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3436..3660
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3706..4134
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3793..3812)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4003..4024
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4143..4163)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4148..4477
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4328..4463
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4390..4409
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4544..5338
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4598..4617)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5208..5227
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5512..5633
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5549..5568)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5603..5622
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5682..5698)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5682..5698)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5695..5717)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5706..5722
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5730..5736,5737..5754,5755..5760))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5775..5796
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5913..5930)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6084..6672)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6164..6183)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6843..7634,7635..7703))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7466..7485
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7704..7808)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
       61 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
      121 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
      181 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
      241 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
      301 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      361 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      421 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      481 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      541 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      601 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      661 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      721 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      781 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
      841 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
      901 acccaagctt ggtaccgagc tcggatccgc caccatgttc gtgtccgatt tccgcaaaga
      961 gttctacgag gtggtccaga gccagagggt ccttctcttc gtggcctcgg acgtggatgc
     1021 tctgtgtgcg tgcaagatcc ttcaggcctt gttccagtgt gaccacgtgc aatatacgct
     1081 ggttccagtt tctgggtggc aagaacttga aactgcattt cttgagcata aagaacagtt
     1141 tcattatttt attctcataa actgtggagc taatgtagac ctattggata ttcttcaacc
     1201 tgatgaagac actatattct ttgtgtgtga cacccatagg ccagtcaatg tcgtcaatgt
     1261 atacaacgat acccagatca aattactcat taaacaagat gatgaccttg aagttcccgc
     1321 ctatgaagac atcttcaggg atgaagagga ggatgaagag cattcaggaa atgacagtga
     1381 tgggtcagag ccttctgaga agcgcacacg gttagaagag gagatagtgg agcaaaccat
     1441 gcggaggagg cagcggcgag agtgggaggc ccggagaaga gacatcctct ttgactacga
     1501 gcagtatgaa tatcatggga catcgtcagc catggtgatg tttgagctgg cttggatgct
     1561 gtccaaggac ctgaatgaca tgctgtggtg ggccatcgtt ggactaacag accagtgggt
     1621 gcaagacaag atcactcaaa tgaaatacgt gactgatgtt ggtgtcctgc agcgccacgt
     1681 ttcccgccac aaccaccgga acgaggatga ggagaacaca ctctccgtgg actgcacacg
     1741 gatctccttt gagtatgacc tccgcctggt gctctaccag cactggtccc tccatgacag
     1801 cctgtgcaac accagctata ccgcagccag gttcaagctg tggtctgtgc atggacagaa
     1861 gcggctccag gagttccttg cagacatggg tcttcccctg aagcaggtga agcagaagtt
     1921 ccaggccatg gacatctcct tgaaggagaa tttgcgggaa atgattgaag agtctgcaaa
     1981 taaatttggg atgaaggaca tgcgcgtgca gactttcagc attcattttg ggttcaagca
     2041 caagtttctg gccagcgacg tggtctttgc caccatgtct ttgatggaga gccccgagaa
     2101 ggatggctca gggacagatc acttcatcca ggctctggac agcctctcca ggagtaacct
     2161 ggacaagctg taccatggcc tggaactcgc caagaagcag ctgcgagcca cccagcagac
     2221 cattgccagc tgcctttgca ccaacctcgt catctcccag gggcctttcc tgtactgctc
     2281 tctcatggag ggcactccag atgtcatgct gttctctagg ccggcatccc taagcctgct
     2341 cagcaaacac ctgctcaagt cctttgtgtg ttcgacaaag aaccggcgct gcaaactgct
     2401 gcccctggtg atggctgccc ccctgagcat ggagcatggc acagtgaccg tggtgggcat
     2461 ccccccagag accgacagct cggacaggaa gaactttttt gggagggcgt ttgagaaggc
     2521 agcggaaagc accagctccc ggatgctgca caaccatttt gacctctcag taattgagct
     2581 gaaagctgag gatcggagca agtttctgga cgcacttatt tccctcctgt ccctcgagca
     2641 tgcatctaga ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     2701 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     2761 gggcgagggc gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa
     2821 gctgcccgtg ccctggccca ccctcgtgac caccctgggc tacggcctgc agtgcttcgc
     2881 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     2941 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     3001 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     3061 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     3121 caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga
     3181 ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3241 cgtgctgctg cccgacaacc actacctgag ctaccagtcc aagctgagca aagaccccaa
     3301 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     3361 catggacgag ctgtacaagt aagggcccta ttctatagtg tcacctaaat gctagagctc
     3421 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     3481 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     3541 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     3601 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     3661 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     3721 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3781 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3841 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3901 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     3961 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4021 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     4081 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     4141 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4201 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4261 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4321 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4381 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     4441 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     4501 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4561 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4621 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4681 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     4741 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4801 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4861 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4921 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4981 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5041 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     5101 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5161 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5221 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5281 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5341 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     5401 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5461 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     5521 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5581 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5641 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     5701 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     5761 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     5821 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5881 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5941 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6001 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6061 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6121 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6181 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6241 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6301 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6361 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6421 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6481 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6541 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6601 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6661 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6721 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6781 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6841 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6901 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6961 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7021 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7081 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7141 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7201 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7261 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7321 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7381 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7441 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7501 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7561 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7621 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7681 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7741 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7801 gttccgcgca catttccccg aaa
//
