LOCUS       Exported                6045 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pmEGFP_a_tubulin_C1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6045)
  AUTHORS   Steigemann P, Wurzenberger C, Schmitz MH, Held M, Guizetti J, Maar 
            S, Gerlich DW
  TITLE     Aurora B-mediated abscission checkpoint protects against 
            tetraploidization.
  JOURNAL   Cell. 2009 Feb 6. 136(3):473-84.
  PUBMED    19203582
REFERENCE   2  (bases 1 to 6045)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6045
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             553..1269
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2773..2894
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2810..2829)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2864..2883
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2901..3356)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3038..3059)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3250..3269
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3383..3487
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3489..3846
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3512..3532)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3697..3832
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3759..3778
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3881..4675
                     /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(3935..3954)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4545..4564
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4863..4882)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4907..4954
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5283..5871
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5772..5791
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tccttaagga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt ccggactcag atctcgagtg cgtgagtgca tctccatcca cgttggccag
     1321 gctggtgtcc agattggcaa tgcctgctgg gagctctact gcctggaaca cggcatccag
     1381 cccgatggcc agatgccaag tgacaagacc attgggggag gagatgactc cttcaacacc
     1441 ttcttcagtg agacgggcgc tggcaagcac gtgccccggg ctgtgtttgt agacttggaa
     1501 cccacagtca ttgatgaagt tcgcactggc acctaccgcc agctcttcca ccctgagcag
     1561 ctcatcacag gcaaggaaga tgctgccaat aactatgccc gagggcacta caccattggc
     1621 aaggagatca ttgaccttgt gttggaccga attcgcaagc tggctgacca gtgcaccggt
     1681 cttcagggct tcttggtttt ccacagcttt ggtgggggaa ctggttctgg gttcacctcc
     1741 ctgctcatgg aacgtctctc agttgattat ggcaagaagt ccaagctgga gttctccatt
     1801 tacccagcac cccaggtttc cacagctgta gttgagccct acaactccat cctcaccacc
     1861 cacaccaccc tggagcactc tgattgtgcc ttcatggtag acaatgaggc catctatgac
     1921 atctgtcgta gaaacctcga tatcgagcgc ccaacctaca ctaaccttaa ccgccttatt
     1981 agccagattg tgtcctccat cactgcttcc ctgagatttg atggagccct gaatgttgac
     2041 ctgacagaat tccagaccaa cctggtgccc tacccccgca tccacttccc tctggccaca
     2101 tatgcccctg tcatctctgc tgagaaagcc taccatgaac agctttctgt agcagagatc
     2161 accaatgctt gctttgagcc agccaaccag atggtgaaat gtgaccctcg ccatggtaaa
     2221 tacatggctt gctgcctgtt gtaccgtggt gacgtggttc ccaaagatgt caatgctgcc
     2281 attgccacca tcaaaaccaa gcgcagcatc cagtttgtgg attggtgccc cactggcttc
     2341 aaggttggca tcaactacca gcctcccact gtggtgcctg gtggagacct ggccaaggta
     2401 cagagagctg tgtgcatgct gagcaacacc acagccattg ctgaggcctg ggctcgcctg
     2461 gaccacaagt ttgacctgat gtatgccaag cgtgcctttg ttcactggta cgtgggtgag
     2521 gggatggagg aaggcgagtt ttcagaggcc cgtgaagata tggctgccct tgagaaggat
     2581 tatgaggagg ttggtgtgga ttctgttgaa ggagagggtg aggaagaagg agaggaatac
     2641 taaggatcca ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt
     2701 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     2761 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2821 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2881 atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     2941 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     3001 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     3061 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     3121 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     3181 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     3241 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     3301 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     3361 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3421 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3481 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     3541 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3601 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3661 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3721 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3781 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3841 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3901 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3961 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4021 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     4081 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4141 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4201 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4261 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4321 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4381 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     4441 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4501 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4561 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4621 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4681 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4741 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4801 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     4861 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     4921 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     4981 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     5041 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     5101 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     5161 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5221 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5281 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5341 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5401 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     5461 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5521 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5581 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5641 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5701 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5761 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5821 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5881 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5941 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
     6001 atcaattacg gggtcattag ttcatagccc atatatggag ttccg
//
