LOCUS       V038845                 7764 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038845
VERSION     V038845
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             874..1590
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             1642..3231
                     /label="ESR2(NM_001437)"
                     /note="ESR2(NM_001437)"
                     /gene="ESR2"
     CDS             3238..3255
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    3331..3555
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3601..4029
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4043..4372
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4439..5233
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5407..5528
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5577..5593)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5601..5617
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5625..5655)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5670..5691
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5979..6564)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6735..7595)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7596..7700)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc
     1621 ggaggcggag ggagtgaatt catggatata aaaaactcac catctagcct taattctcct
     1681 tcctcctaca actgcagtca atccatctta cccctggagc acggctccat atacatacct
     1741 tcctcctatg tagacagcca ccatgaatat ccagccatga cattctatag ccctgctgtg
     1801 atgaattaca gcattcccag caatgtcact aacttggaag gtgggcctgg tcggcagacc
     1861 acaagcccaa atgtgttgtg gccaacacct gggcaccttt ctcctttagt ggtccatcgc
     1921 cagttatcac atctgtatgc ggaacctcaa aagagtccct ggtgtgaagc aagatcgcta
     1981 gaacacacct tacctgtaaa cagagagaca ctgaaaagga aggttagtgg gaaccgttgc
     2041 gccagccctg ttactggtcc aggttcaaag agggatgctc acttctgcgc tgtctgcagc
     2101 gattacgcat cgggatatca ctatggagtc tggtcgtgtg aaggatgtaa ggcctttttt
     2161 aaaagaagca ttcaaggaca taatgattat atttgtccag ctacaaatca gtgtacaatc
     2221 gataaaaacc ggcgcaagag ctgccaggcc tgccgacttc ggaagtgtta cgaagtggga
     2281 atggtgaagt gtggctcccg gagagagaga tgtgggtacc gccttgtgcg gagacagaga
     2341 agtgccgacg agcagctgca ctgtgccggc aaggccaaga gaagtggcgg ccacgcgccc
     2401 cgagtgcggg agctgctgct ggacgccctg agccccgagc agctagtgct caccctcctg
     2461 gaggctgagc cgccccatgt gctgatcagc cgccccagtg cgcccttcac cgaggcctcc
     2521 atgatgatgt ccctgaccaa gttggccgac aaggagttgg tacacatgat cagctgggcc
     2581 aagaagattc ccggctttgt ggagctcagc ctgttcgacc aagtgcggct cttggagagc
     2641 tgttggatgg aggtgttaat gatggggctg atgtggcgct caattgacca ccccggcaag
     2701 ctcatctttg ctccagatct tgttctggac agggatgagg ggaaatgcgt agaaggaatt
     2761 ctggaaatct ttgacatgct cctggcaact acttcaaggt ttcgagagtt aaaactccaa
     2821 cacaaagaat atctctgtgt caaggccatg atcctgctca attccagtat gtaccctctg
     2881 gtcacagcga cccaggatgc tgacagcagc cggaagctgg ctcacttgct gaacgccgtg
     2941 accgatgctt tggtttgggt gattgccaag agcggcatct cctcccagca gcaatccatg
     3001 cgcctggcta acctcctgat gctcctgtcc cacgtcaggc atgcgagtaa caagggcatg
     3061 gaacatctgc tcaacatgaa gtgcaaaaat gtggtcccag tgtatgacct gctgctggag
     3121 atgctgaatg cccacgtgct tcgcgggtgc aagtcctcca tcacggggtc cgagtgcagc
     3181 ccggcagagg acagtaaaag caaagagggc tcccagaacc cacagtctca gggtacccat
     3241 caccatcatc accattagga tcctgatatc cagcacagtg gcggccgctc gagtctagag
     3301 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     3361 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     3421 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     3481 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     3541 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     3601 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3661 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3721 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3781 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3841 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3901 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3961 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4021 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4081 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4141 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4201 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4261 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     4321 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     4381 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     4441 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4501 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4561 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4621 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4681 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4741 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4801 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4861 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4921 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4981 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5041 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5101 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5161 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5221 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5281 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5341 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5401 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5461 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5521 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     5581 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5641 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5701 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5761 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5821 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5881 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5941 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6001 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6061 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6121 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6181 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6241 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6301 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6361 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6421 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6481 tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt
     6541 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6601 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6661 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6721 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6781 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6841 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6901 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6961 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7021 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7081 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7141 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7201 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7261 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7321 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7381 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7441 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7501 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7561 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7621 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7681 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     7741 ggagatctcc cgatccccta tggt
//
