LOCUS       sequence-350799-        5661 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 5661)
  AUTHORS   Wang W, Budhu A, Forgues M, Wang XW
  TITLE     Temporal and spatial control of nucleophosmin by the Ran-Crm1 
            complex in centrosome duplication.
  JOURNAL   Nat Cell Biol. 2005 Aug . 7(8):823-30.
  PUBMED    16041368
REFERENCE   2  (bases 1 to 5661)
  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: "Nat Cell 
            Biol. 2005 Aug . 7(8):823-30."
COMMENT     Sequence Label: GFP-NPM WT
FEATURES             Location/Qualifiers
     source          1..5661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        101..292
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     249..269
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(294..398)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      425..880
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(512..531)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     722..743
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    887..1008
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(898..917)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     952..971
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2026..2049)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      2049..2058
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2062..2127
                     /label=MCS
                     /note="multiple cloning site"
     CDS             complement(join(2128..2838,2839..2841,2842..2844))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(2128..2838,2839..2841,2842..2844))
                     /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(2170..2191)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2519..2538
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2778..2799
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2841..2850
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2889..3092)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2918..2938)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3093..3396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3571..4159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3651..3670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4488..4535
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4560..4579
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4767..5561)
                     /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(4767..5561)
                     /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(4878..4897)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5488..5507
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5596..5661,1..292))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5610..5661,1..84)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
       61 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
      121 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
      181 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
      241 ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc aggactcttc
      301 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
      361 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
      421 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
      481 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
      541 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
      601 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
      661 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
      721 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
      781 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
      841 tttaacgcga attttaacaa aatattaacg cttacaattt acgcgttaag atacattgat
      901 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
      961 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     1021 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     1081 aacctctaca aatgtggtat ggctgattat gatcagttat ctagatccgg tggatcctta
     1141 aagagacttc ctccactgcc agagatcttg aatagcctct tggtcagtca tccggaagca
     1201 attcttcaca taattgatga atttggcttc cactttggga agagaaccac ctttttctat
     1261 acttgcttgc atttttgctt taatgtcttc tacagaacta ggtccttttg gtgttttagg
     1321 agttttttcc tgtttcttga aggattcttg tccttttgat cttggtgttg atgatggttt
     1381 tgagtctttt ccattctgat ttgacttttg tgcatttttg gctggagtat ctcgtataga
     1441 tttcttcact ggcgcttttt cttcagcttc ctcatcatca aaatcatcat catcatcatc
     1501 ttcatcatca tcctcttcat catcatcgtc atcatcttca tcagcagcaa gttttacttt
     1561 tttctgtgga accttgctac cacctccagg ggcagaccgc tttccagata tacttaagag
     1621 tttcacatcc tcctcctctt catcttctga ctctgcatct tcctccacag ctactaagtg
     1681 ctgtccacta atatgcactg gccctgaacc acacttcaac cttaagacca ctggtggtgt
     1741 tatttcaaag cccccaaggg aaaccgttgg ctgtacagac attttcaaag ttgccagtgt
     1801 tactttaatt ggactgcctt cgtaattcat tgcctctgct tcaacaatgt gcaactcatc
     1861 ctttgcacca gcccctaaac tgaccgttct taaagataac tggtgctcat tttcatcatt
     1921 atccacctta aagtgataat ctttgtcggc ctttagttca caaccgaaaa gatagttctg
     1981 gggcctcagg gggctcatgt ccatgtccat cgaatcttcc atatgcttgt cgtcgtcatc
     2041 cttgtagtcc atggtggcgg cggatcccgg gcccgcggta ccgtcgactg cagaattcga
     2101 agcttgagct cgagatctga gtccggactt gtacagctcg tccatgccga gagtgatccc
     2161 ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct
     2221 cagggcggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat
     2281 gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg
     2341 gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg gccatgatat agacgttgtg
     2401 gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat
     2461 gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt
     2521 cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat
     2581 ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac
     2641 gccgtaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca
     2701 gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct
     2761 gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa
     2821 cagctcctcg cccttgctca ccatggtggc gaccggtagc gctagcggat ctgacggttc
     2881 actaaaccag ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc
     2941 aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa
     3001 ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca
     3061 cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag
     3121 atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc
     3181 catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact
     3241 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt
     3301 ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt
     3361 cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga
     3421 actaatgacc ccgtaattga ttactattaa taactaatgc atggcggtaa tacggttatc
     3481 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3541 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3601 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3661 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3721 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3781 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3841 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3901 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3961 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4021 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4081 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4141 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4201 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4261 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaacctga
     4321 ggctatggca gggcctgccg ccccgacgtt ggctgcgagc cctgggcctt cacccgaact
     4381 tggggggtgg ggtggggaaa aggaagaaac gcgggcgtat tggccccaat ggggtctcgg
     4441 tggggtatcg acagagtgcc agccctggga ccgaaccccg cgtttatgaa caaacgaccc
     4501 aacaccgtgc gttttattct gtctttttat tgccgtcata gcgcgggttc cttccggtat
     4561 tgtctccttc cgtgtttcag ttagcctccc cctagggtgg gcgaagaact ccagcatgag
     4621 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
     4681 ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc
     4741 ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg
     4801 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
     4861 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc
     4921 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
     4981 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg
     5041 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     5101 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     5161 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     5221 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     5281 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     5341 cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg
     5401 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     5461 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     5521 gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct
     5581 gtctcttgat cgatctttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     5641 gaatagctca gaggccgagg c
//
