LOCUS       sequence-416906-        5076 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GFP-tagged hsRPA3 in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5076)
  AUTHORS   Haring SJ, Mason AC, Binz SK, Wold MS
  TITLE     Cellular functions of human RPA1. Multiple roles of domains in 
            replication, repair, and checkpoints.
  JOURNAL   J Biol Chem. 2008 Jul 4;283(27):19095-111. doi: 
            10.1074/jbc.M800881200. Epub 2008 May 9.
  PUBMED    18469000
REFERENCE   2  (bases 1 to 5076)
  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"; doi: 
            "10.1074/jbc.M800881200"; journalName: "J Biol Chem"; date: 
            "2008-07-4- 4"; volume: "283"; issue: "27"; pages: "19095-111"
COMMENT     Sequence Label: pEGFP-hsRPA14 (MSW#503)
FEATURES             Location/Qualifiers
     source          1..5076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(351..1061,1062..1064,1065..1067))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(351..1061,1062..1064,1065..1067))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(393..414)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     742..761
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1001..1022
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1064..1073
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1112..1315)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1141..1161)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1316..1619
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1794..2382)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1874..1893)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2711..2758
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2783..2802
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2990..3784)
                     /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             complement(2990..3784)
                     /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(3101..3120)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3711..3730
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3819..4176)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3833..3968
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3985..4176
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4133..4153
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4178..4282)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4309..4764
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4396..4415)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4606..4627
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4771..4892
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4782..4801)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4836..4855
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 aagtcatgga taattttcac agcttcattg taaagtccaa gatcaaaagg atggctatct
       61 tctttaaact ggacataaga tgtacacaag atggtggcct tggcggttac tcttccaacc
      121 acttccacaa ttccagagat ttcttcatca aggggttcca tcaactcgat ggttccattt
      181 tttccttctc catctgaaag aataaacatt tttccggtgg gatgaatctt ttccagcctc
      241 cctacgaagc agacaggctt gtcgatgaat tgagctagca tgccggcgtt gatgcgcgac
      301 ctgggcaagt ccatcatgtc caccatgcct ccaccagatc tgagtccgga cttgtacagc
      361 tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg
      421 cgcttctcgt tggggtcttt gctcagggcg gactgggtgc tcaggtagtg gttgtcgggc
      481 agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg
      541 ctgccgtcct cgatgttgtg gcggatcttg aagttcacct tgatgccgtt cttctgcttg
      601 tcggccatga tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg
      661 ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc
      721 tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc
      781 tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg
      841 gggtagcggc tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc
      901 acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg
      961 ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc
     1021 aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt
     1081 agcgctagcg gatctgacgg ttcactaaac cagctctgct tatatagacc tcccaccgta
     1141 cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc
     1201 gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc
     1261 cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg
     1321 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     1381 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     1441 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     1501 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     1561 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     1621 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat taataactaa
     1681 tgcatggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1741 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1801 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1861 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1921 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     1981 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2041 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2101 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2161 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2221 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2281 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2341 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2401 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2461 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2521 taaagtatat atgagtaacc tgaggctatg gcagggcctg ccgccccgac gttggctgcg
     2581 agccctgggc cttcacccga acttgggggg tggggtgggg aaaaggaaga aacgcgggcg
     2641 tattggcccc aatggggtct cggtggggta tcgacagagt gccagccctg ggaccgaacc
     2701 ccgcgtttat gaacaaacga cccaacaccg tgcgttttat tctgtctttt tattgccgtc
     2761 atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct ccccctaggg
     2821 tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg
     2881 aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtgat
     2941 ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct cagaagaact
     3001 cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca
     3061 cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg
     3121 ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc
     3181 ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct
     3241 cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat
     3301 gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct
     3361 cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc
     3421 gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga
     3481 gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt
     3541 cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt
     3601 cttgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct
     3661 gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat
     3721 agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa
     3781 tcatgcgaaa cgatcctcat cctgtctctt gatcgatctt tgcaaaagcc taggcctcca
     3841 aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca
     3901 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
     3961 ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt
     4021 tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg
     4081 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac
     4141 tgacacacat tccacagctg gttctttccg cctcaggact cttccttttt caatattatt
     4201 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4261 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta
     4321 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4381 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4441 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4501 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4561 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4621 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4681 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4741 acaaaatatt aacgcttaca atttacgcgt taagatacat tgatgagttt ggacaaacca
     4801 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     4861 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     4921 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     4981 gtatggctga ttatgatcag ttatctagat ccggtggatc ccgggcccgc ggtacctcaa
     5041 tcatgttgca caatccctaa aggataaaac tgaggg
//
