LOCUS       Exported                7162 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pEGFP-D50 lamin A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7162)
  AUTHORS   Scaffidi P, Misteli T
  TITLE     Reversal of the cellular phenotype in the premature aging disease 
            Hutchinson-Gilford progeria syndrome.
  JOURNAL   Nat Med. 2005 Apr . 11(4):440-5.
  PUBMED    15750600
REFERENCE   2  (bases 1 to 7162)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7162
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             613..1329
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             613..1329
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3950..4071
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3987..4006)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4041..4060
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4078..4533)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4215..4236)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4427..4446
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4560..4664
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4666..5023
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4689..4709)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4874..5009
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4936..4955
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5058..5852
                     /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(5112..5131)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5722..5741
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6040..6059)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6084..6131
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6460..7048
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6949..6968
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc
     1381 gcgggcagcg ctgccaacct gccggccatg gagaccccgt cccagcggcg cgccacccgc
     1441 agcggggcgc aggccagctc cactccgctg tcgcccaccc gcatcacccg gctgcaggag
     1501 aaggaggacc tgcaggagct caatgatcgc ttggcggtct acatcgaccg tgtgcgctcg
     1561 ctggaaacgg agaacgcagg gctgcgcctt cgcatcaccg agtctgaaga ggtggtcagc
     1621 cgcgaggtgt ccggcatcaa ggccgcctac gaggccgagc tcggggatgc ccgcaagacc
     1681 cttgactcag tagccaagga gcgcgcccgc ctgcagctgg agctgagcaa agtgcgtgag
     1741 gagtttaagg agctgaaagc gcgcaatacc aagaaggagg gtgacctgat agctgctcag
     1801 gctcggctga aggacctgga ggctctgctg aactccaagg aggccgcact gagcactgct
     1861 ctcagtgaga agcgcacgct ggagggcgag ctgcatgatc tgcggggcca ggtggccaag
     1921 cttgaggcag ccctaggtga ggccaagaag caacttcagg atgagatgct gcggcgggtg
     1981 gatgctgaga acaggctgca gaccatgaag gaggaactgg acttccagaa gaacatctac
     2041 agtgaggagc tgcgtgagac caagcgccgt catgagaccc gactggtgga gattgacaat
     2101 gggaagcagc gtgagtttga gagccggctg gcggatgcgc tgcaggaact gcgggcccag
     2161 catgaggacc aggtggagca gtataagaag gagctggaga agacttattc tgccaagctg
     2221 gacaatgcca ggcagtctgc tgagaggaac agcaacctgg tgggggctgc ccacgaggag
     2281 ctgcagcagt cgcgcatccg catcgacagc ctctctgccc agctcagcca gctccagaag
     2341 cagctggcag ccaaggaggc gaagcttcga gacctggagg actcactggc ccgtgagcgg
     2401 gacaccagcc ggcggctgct ggcggaaaag gagcgggaga tggccgagat gcgggcaagg
     2461 atgcagcagc agctggacga gtaccaggag cttctggaca tcaagctggc cctggacatg
     2521 gagatccacg cctaccgcaa gctcttggag ggcgaggagg agaggctacg cctgtccccc
     2581 agccctacct cgcagcgcag ccgtggccgt gcttcctctc actcatccca gacacagggt
     2641 gggggcagcg tcaccaaaaa gcgcaaactg gagtccactg agagccgcag cagcttctca
     2701 cagcacgcac gcactagcgg gcgcgtggcc gtggaggagg tggatgagga gggcaagttt
     2761 gtccggctgc gcaacaagtc caatgaggac cagtccatgg gcaattggca gatcaagcgc
     2821 cagaatggag atgatccctt gctgacttac cggttcccac caaagttcac cctgaaggct
     2881 gggcaggtgg tgacgatctg ggctgcagga gctggggcca cccacagccc ccctaccgac
     2941 ctggtgtgga aggcacagaa cacctggggc tgcgggaaca gcctgcgtac ggctctcatc
     3001 aactccactg gggaagaagt ggccatgcgc aagctggtgc gctcagtgac tgtggttgag
     3061 gacgacgagg atgaggatgg agatgacctg ctccatcacc accacggctc ccactgcagc
     3121 agctcggggg accccgctga gtacaacctg cgctcgcgca ccgtgctgtg cgggacctgc
     3181 gggcagcctg ccgacaaggc atctgccagc ggctcaggag cccagagccc ccagaactgc
     3241 agcatcatgt aatctgggac ctgccaggca ggggtggggg tggaggcttc ctgcgtcctc
     3301 ctcacctcat gcccaccccc tgccctgcac gtcatgggag ggggcttgaa gccaaagaaa
     3361 aataaccctt tggttttttt cttctgtatt tttttttcta agagaagtta ttttctacag
     3421 tggttttata ctgaaggaaa aacacaagca aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     3481 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     3541 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     3601 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     3661 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     3721 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     3781 aaaaaaaaag cgtctagaaa ccctttagat ctgctgcagc ggatccaccg gatctagata
     3841 actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     3901 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     3961 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4021 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa
     4081 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     4141 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     4201 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     4261 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     4321 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     4381 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     4441 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     4501 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     4561 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4621 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     4681 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4741 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4801 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4861 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4921 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4981 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     5041 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     5101 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     5161 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     5221 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     5281 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     5341 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     5401 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5461 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5521 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5581 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5641 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5701 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5761 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5821 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5881 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5941 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     6001 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     6061 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     6121 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     6181 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     6241 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     6301 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6361 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6421 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6481 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6541 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6601 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6661 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6721 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6781 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6841 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6901 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6961 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     7021 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     7081 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     7141 taaccgtatt accgccatgc at
//
