LOCUS       Exported                5963 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-PV-NES-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5963)
  AUTHORS   Pusl T, Wu JJ, Zimmerman TL, Zhang L, Ehrlich BE, Berchtold MW, Hoek
            JB, Karpen SJ, Nathanson MH, Bennett AM
  TITLE     Epidermal growth factor-mediated activation of the ETS domain 
            transcription factor Elk-1 requires nuclear calcium.
  JOURNAL   J Biol Chem. 2002 Jul 26. 277(30):27517-27.
  PUBMED    11971908
REFERENCE   2  (bases 1 to 5963)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5963
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        12..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      618..627
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1029..1745
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1062..1090)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1685..1705
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1755..1784
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1820..1837)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1826..2050
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2096..2524
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2183..2202)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2393..2414
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2533..2553)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2538..2867
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2718..2853
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2780..2799
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2893..3687
                     /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="MVEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2947..2966)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3557..3576
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3861..3982
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3898..3917)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3952..3971
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4015..4032)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4186..4774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4266..4285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4945..5805)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5568..5587
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5806..5910)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtaccgaatt cacattgatt attgagtagt tattaatagt aatcaattac ggggtcatta
       61 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      121 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      181 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      241 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      301 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      361 atctacggtt agtcatcgct attaccatag tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctttctggc
      601 taactagaga acccgtggcc accatggccc aggtgcagct gcaggtcgac gcattacaaa
      661 aaaaattaga agaattagaa ttagatgaaa tgtcgatgac agacttgctc agcgctgagg
      721 acatcaagaa ggcgatagga gcctttactg ctgcagactc cttcgaccac aaaaagttct
      781 tccagatggt gggcctgaag aaaaagagtg cggatgatgt gaagaaggtg ttccacattc
      841 tggacaaaga caaaagtggc ttcattgagg aggatgagct ggggtccatt ctgaagggct
      901 tctcctcaga tgccagagac ttgtctgcta aggaaacaaa gacgctgatg gctgctggag
      961 acaaggacgg ggacggcaag attggggttg aagaattctc cactctggtg gccgaaagcg
     1021 cggccgcaat ggctagcaaa ggagaagaac ttttcactgg agttgtccca attcttgttg
     1081 aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg
     1141 ctacatacgg aaagcttacc cttaaattta tttgcactac tggaaaacta cctgttccat
     1201 ggccaacact tgtcactact ttctcttatg gtgttcaatg cttttcccgt tatccggatc
     1261 atatgaaacg gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaacgca
     1321 ctatatcttt caaagatgac gggaactaca agacgcgtgc tgaagtcaag tttgaaggtg
     1381 atacccttgt taatcgtatc gagttaaaag gtattgattt taaagaagat ggaaacattc
     1441 tcggacacaa actcgagtac aactataact cacacaatgt atacatcacg gcagacaaac
     1501 aaaagaatgg aatcaaagct aacttcaaaa ttcgccacaa cattgaagat ggatccgttc
     1561 aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag
     1621 acaaccatta cctgtcgaca caatctgccc tttcgaaaga tcccaacgaa aagcgtgacc
     1681 acatggtcct tcttgagttt gtaactgctg ctgggattac acatggcatg gatgagctct
     1741 acaaagcggc cgcagaacaa aaactcatct cagaagagga tctgaatggg gccgcatagt
     1801 ctagaagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     1861 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     1921 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     1981 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatggcccg
     2041 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     2101 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2161 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2221 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2281 ttacggcacc tctcccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     2341 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2401 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     2461 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2521 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2581 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     2641 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     2701 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2761 taggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     2821 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccccccc
     2881 gggaggtcca caatggttga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     2941 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3001 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3061 ctgaatgaac tccaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3121 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3181 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3241 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3301 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3361 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3421 cgtatgcccg acggcgagga tctcgtcgtg actcatggcg atgcctgctt gccgaatatc
     3481 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3541 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     3601 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     3661 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     3721 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     3781 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     3841 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     3901 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3961 ccaaactcat caatgtatct tatcatgtct gtataccgtc gatctttccg cttcctcgct
     4021 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4081 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4141 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4201 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4261 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4321 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4381 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtat
     4441 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4501 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4561 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4621 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4681 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4741 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4801 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4861 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4921 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4981 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5041 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5101 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5161 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5221 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5281 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5341 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5401 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5461 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5521 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5581 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5641 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgggcac ccaactgatc
     5701 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5761 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5821 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5881 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     5941 cagatcgacg gatcgggaga tcg
//
