LOCUS       Exported                7231 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    GFP-UBF
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7231)
  AUTHORS   Dundr M, Hoffmann-Rohrer U, Hu Q, Grummt I, Rothblum LI, Phair RD, 
            Misteli T
  TITLE     A kinetic framework for a mammalian RNA polymerase in vivo.
  JOURNAL   Science. 2002 Nov 22. 298(5598):1623-6.
  PUBMED    12446911
REFERENCE   2  (bases 1 to 7231)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7231
                     /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"
     protein_bind    3385..3396
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     polyA_signal    4019..4140
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4056..4075)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4110..4129
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4147..4602)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4284..4305)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4496..4515
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4629..4733
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4735..5092
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4758..4778)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4943..5078
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5005..5024
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5127..5921
                     /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(5181..5200)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5791..5810
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6109..6128)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6153..6200
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6529..7117
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7018..7037
                     /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 ccggtatgaa cggagaagcc gactgcccca cagacctgga aatggccgcc
     1381 cccaaaggcc aagaccgttg gtcccaggaa gacatgctga ctttgctgga atgcatgaag
     1441 aacaaccttc catccaatga cagctccaag ttcaaaacca ccgaatcaca catggactgg
     1501 gaaaaagtag catttaaaga cttttctgga gacatgtgca agctcaaatg ggtggagatt
     1561 tctaatgagg tgaggaagtt ccgtacattg acagaattga tcctcgatgc tcaggaacat
     1621 gttaaaaatc cttacaaagg caaaaaactc aagaaacacc cagacttccc aaagaagccc
     1681 ctgacccctt atttccgctt cttcatggag aagcgggcca agtatgcgaa actccaccct
     1741 gagatgagca acctggacct aaccaagatt ctgtccaaga aatacaagga gcttccggag
     1801 aagaagaaga tgaaatatat tcaggacttc cagagagaga aacaggagtt cgagcgaaac
     1861 ctggcccgat tcagggagga tcaccccgac ctaatccaga atgccaagaa atcggacatc
     1921 ccagagaagc ccaaaacccc ccagcagctg tggtacaccc acgagaagaa ggtgtatctc
     1981 aaagtgcggc cagatgccac tacgaaggag gtgaaggact ccctggggaa gcagtggtct
     2041 cagctctcgg acaaaaagag gctgaaatgg attcataagg ccctggagca gcggaaggag
     2101 tacgaggaga tcatgagaga ctatatccag aagcacccag agctgaacat cagtgaggag
     2161 ggtatcacca agtccaccct caccaaggcc gaacgccagc tcaaggacaa gtttgacggg
     2221 cgacccacca agccacctcc gaacagctac tcgctgtact gcgcagagct catggccaac
     2281 atgaaggacg tgcccagcac agagcgcatg gtgctgtgca gccagcagtg gaagctgctg
     2341 tcccagaagg agaaggacgc ctatcacaag aagtgtgatc agaaaaagaa agattacgag
     2401 gtggagctgc tccgtttcct cgagagcctg cctgaggagg agcagcagcg ggtcttgggg
     2461 gaagagaaga tgctgaacat caacaagaag caggccacca gccccgcctc caagaagcca
     2521 gcccaggaag ggggcaaggg cggctccgag aagcccaagc ggcccgtgtc ggccatgttc
     2581 atcttctcgg aggagaaacg gcggcagctg caggaggagc ggcctgagct ctccgagagc
     2641 gagctgaccc gcctgctggc ccgaatgtgg aacgacctgt ctgagaagaa gaaggccaag
     2701 tacaaggccc gagaggcggc gctcaaggct cagtcggaga ggaagcccgg cggggagcgc
     2761 gaggaacggg gcaagctgcc cgagtccccc aaaagagctg aggagatctg gcaacagagc
     2821 gttatcggcg actacctggc ccgcttcaag aatgaccggg tgaaggcctt gaaagccatg
     2881 gaaatgacct ggaataacat ggaaaagaag gagaaactga tgtggattaa gaaggcagcc
     2941 gaagaccaaa agcgatatga gagagagctg agtgagatgc gggcacctcc agctgctaca
     3001 aattcttcca agaagatgaa attccaggga gaacccaaga agcctcccat gaacggttac
     3061 cagaagttct cccaggagct gctgtccaat ggggagctga accacctgcc gctgaaggag
     3121 cgcatggtgg agatcggcag tcgctggcag cgcatctccc agagccagaa ggagcactac
     3181 aaaaagctgg ccgaggagca gcaaaagcag tacaaggtgc acctggacct ctgggttaag
     3241 agcctgtctc cccaggaccg tgcagcatat aaagagtaca tctccaataa acgtaagagc
     3301 atgaccaagc tgcgaggccc aaaccccaaa tccagccgga ctactctgca gtccaagtcg
     3361 gagtccgagg aggatgatga agaggatgag gatgacgagg acgaggatga agaagaggaa
     3421 gatgatgaga atggggactc ctctgaagat ggcggcgact cctctgagtc cagcagcgag
     3481 gacgagagcg aggatgggga tgagaatgaa gaggatgacg aggacgaaga cgacgacgag
     3541 gatgacgatg aggatgaaga taatgagtcc gagggcagca gctccagctc ctcctcctca
     3601 ggggactcct cagactctga ctccaactga ggctcagccc caccccaggg cagccaggga
     3661 gagcccagga gctcccctcc ccaactgacc acctttgttt ctcccccatg ttctgtccct
     3721 tgcccccctg gcctccccca ctttctttct ttctttaaaa aaaaaaaaaa aaatacggtg
     3781 ggggtagggg gctggaggag cccaggccag gactctgcag cctcagagac atcagccctt
     3841 gggggtcctc ctccagggac agcaactatc agactaagcc agcaccggag gatccaccgg
     3901 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     3961 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     4021 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4081 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4141 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     4201 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     4261 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     4321 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     4381 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     4441 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     4501 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4561 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     4621 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4681 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     4741 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4801 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4861 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4921 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4981 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     5041 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     5101 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     5161 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     5221 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     5281 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     5341 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     5401 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     5461 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     5521 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5581 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5641 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5701 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5761 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5821 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     5881 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     5941 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     6001 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     6061 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     6121 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     6181 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     6241 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     6301 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     6361 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6421 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6481 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6541 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6601 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6661 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6721 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6781 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6841 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6901 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6961 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7021 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7081 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7141 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     7201 ttctgtggat aaccgtatta ccgccatgca t
//
