LOCUS       sequence-327709-        6641 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6641)
  AUTHORS   Joseph J, Tan SH, Karpova TS, McNally JG, Dasso M
  TITLE     SUMO-1 targets RanGAP1 to kinetochores and mitotic spindles.
  JOURNAL   J Cell Biol. 2002 Feb 18. 156(4):595-602.
  PUBMED    11854305
REFERENCE   2  (bases 1 to 6641)
  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: "J Cell 
            Biol. 2002 Feb 18. 156(4):595-602."
COMMENT     Sequence Label: pEGFP-C2 RanGAP
FEATURES             Location/Qualifiers
     source          1..6641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      243..252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(249..251,252..254,255..965)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(249..251,252..254,255..965)
                     /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(555..574)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1955..1972
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     CDS             2859..2900
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2910..2927
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3065..3186
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3102..3121)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3156..3175
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3193..3648)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3330..3351)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3542..3561
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3675..3779
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3781..4138
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3781..3972
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3804..3824)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3989..4124
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4173..4967
                     /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             4173..4967
                     /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(4227..4246)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4837..4856
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5155..5174)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5199..5246
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5575..6163
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6064..6083
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6338..6641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      301 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      361 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      421 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
      481 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      541 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      601 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      661 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
      721 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
      781 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      841 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
      901 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
      961 acaagtccgg ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc
     1021 gcgggcccgg gatggcctcg gaagacattg ccaagctggc agagacactt gccaagactc
     1081 aggtggccgg gggacagctg agtttcaaag gcaagagcct caaactcaac actgcagaag
     1141 atgctaaaga tgtgattaaa gagattgaag actttgacag cttggaggct ctgcgtctgg
     1201 aaggcaacac agtgggcgtg gaagccgcca gggtcatcgc caaggcctta gagaagaagt
     1261 cggagttgaa gcgctgccac tggagtgaca tgttcacggg aaggctgcgg accgagatcc
     1321 caccagccct gatctcacta ggggaaggac tcatcacagc tggggctcag ctggtggagc
     1381 tggacttaag cgacaacgca ttcgggcccg acggtgtgca aggcttcgag gccctgctca
     1441 agagctcagc ctgcttcacc ctgcaggaac tcaagctcaa caactgtggc atgggcattg
     1501 gcggcggcaa gatcctggct gcagctctga ccgaatgtca ccggaaatcc agtgcccaag
     1561 gcaagcctct ggccctgaag gtctttgtgg ctggcagaaa ccgtctggag aatgatggcg
     1621 ccactgcctt ggcagaagct tttagggtca tcgggaccct ggaggaggtc cacatgccac
     1681 agaatgggat caaccaccct ggcatcactg ccctggccca ggctttcgct gtcaaccccc
     1741 tgctgcgggt catcaacctg aatgacaaca ccttcactga gaagggcgcc gtggccatgg
     1801 ccgagacctt gaagaccttg cggcaggtgg aggtgattaa ttttggggac tgcctggtgc
     1861 gctccaaggg tgcagttgcc attgcagatg ccatccgcgg cggcctgccc aagctaaagg
     1921 agctgaactt gtcattctgt gagatcaaga gggatgctgc cctggctgtt gctgaggcca
     1981 tggcagacaa agctgagctg gagaagctgg acctgaatgg caacaccctg ggagaagaag
     2041 gctgtgaaca gcttcaggag gtgctggagg gcttcaacat ggccaaggtg ctggcgtccc
     2101 tcagtgatga cgaggacgag gaggaggagg aggaaggaga agaggaagaa gaggaagcag
     2161 aagaagagga ggaggaagat gaggaagagg aggaagaaga ggaggaggag gaggaagaag
     2221 agcctcagca gcgagggcag ggagagaagt cagccacgcc ctcacggaag attctggacc
     2281 ctaacactgg ggagccagct cccgtgctgt cctccccacc tcctgcagac gtctccacct
     2341 tcctggcttt tccctctcca gagaagctgc tgcgcctagg gcccaagagc tccgtgctga
     2401 tagcccagca gactgacacg tctgaccccg agaaggtggt ctctgccttc ctaaaggtgt
     2461 catctgtgtt caaggacgaa gctactgtga ggatggcagt gcaggatgca gtagatgccc
     2521 tgatgcagaa ggctttcaac tcctcgtcct tcaactccaa caccttcctc accaggctcc
     2581 tcgtgcacat gggtctgctc aagagtgaag acaaggtcaa ggccattgcc aacctgtacg
     2641 gccccctgat ggcgctgaac cacatggtgc agcaggacta tttccccaag gcccttgcac
     2701 ccctgctgct ggcgttcgtg accaagccca acagcgccct ggaatcctgc tccttcgccc
     2761 gccacagtct gctgcagacg ctgtacaagg tcaagggcaa ttctgcagat atccagcaca
     2821 gtggcggccg ctcgagtcta gagggcccgc ggttcgaagg taagcctatc cctaaccctc
     2881 tcctcggtct cgattctacg cgtaccggtc atcatcacca tcaccattga gtttgggatc
     2941 caccggatct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
     3001 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3061 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3121 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3181 atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3241 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3301 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3361 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3421 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3481 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3541 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3601 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     3661 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3721 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     3781 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3841 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3901 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3961 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4021 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4081 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     4141 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4201 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4261 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4321 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     4381 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4441 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4501 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4561 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4621 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4681 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4741 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4801 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4861 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4921 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4981 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5041 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5101 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     5161 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     5221 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     5281 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     5341 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     5401 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     5461 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5521 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5581 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5641 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5701 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5761 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5821 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5881 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5941 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6001 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6061 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6121 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6181 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6241 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
     6301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     6361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     6421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     6481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     6541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     6601 cttggcagta catctacgta ttagtcatcg ctattaccat g
//
