LOCUS       V040073                 8467 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040073
VERSION     V040073
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..3957
                     /label="NFE2L1(NM_003204)"
                     /note="NFE2L1(NM_003204)"
                     /gene="NFE2L1"
     polyA_signal    4034..4258
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4304..4732
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4746..5075
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5142..5936
                     /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    6110..6231
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6280..6296)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6304..6320
                     /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(6328..6358)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6373..6394
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6682..7267)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7438..8298)
                     /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(8299..8403)
                     /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 catgctttct ctgaagaaat acttaacgga aggacttctc
     1681 cagttcacca ttctgctgag tttgattggg gtacgggtgg acgtggatac ttacctgacc
     1741 tcacagcttc ccccactccg ggagatcatc ctggggccca gttctgccta tactcagacc
     1801 cagttccaca acctgaggaa taccttggat ggctatggta tccaccccaa gagcatagac
     1861 ctggacaatt acttcactgc ccggcggctc ctcagtcagg tgagggccct ggacaggttc
     1921 caggtgccaa ccactgaggt aaatgcctgg ctggttcacc gagacccaga ggggtctgtc
     1981 tctggcagtc agcccaactc aggcctcgcc ctcgagagtt ccagtggcct ccaagatgtg
     2041 acaggcccag acaacggggt gcgagaaagc gaaacggagc agggattcgg tgaagatttg
     2101 gaggatttgg gggctgtagc ccccccagtc agtggagact taaccaaaga ggacatagat
     2161 ctgattgaca tcctttggcg acaggatatt gatctggggg ctgggcgtga ggtttttgac
     2221 tatagtcacc gccagaagga gcaggatgtg gagaaggagc tgcgagatgg aggcgagcag
     2281 gacacctggg caggcgaggg cgcggaagct ctggcacgga acctgctagt ggatggagag
     2341 actggggaga gcttccctgc acaggtgcct agtggggagg accagacggc cctgtccctg
     2401 gaagagtgcc ttaggctgct ggaagccacc tgcccctttg gggagaatgc tgagtttcca
     2461 gcagacattt ccagcataac agaagcagtg cctagtgaga gtgagccccc tgctcttcaa
     2521 aacaacctct tgtctcctct tctgaccggg acagagtcac catttgattt ggaacagcag
     2581 tggcaagatc tcatgtccat catggaaatg caggccatgg aagtgaacac atcagcaagt
     2641 gaaatcctgt acagtgcccc tcctggagac ccactgagca ccaactacag ccttgccccc
     2701 aacactccca tcaatcagaa tgtcagcctg catcaggcgt ccctgggggg ctgcagccag
     2761 gacttcttac tcttcagccc cgaggtggaa agcctgcctg tggccagtag ctccacgctg
     2821 ctcccgttgg cccccagcaa ttctaccagc ctcaactcca ccttcggctc caccaacctg
     2881 acagggctct tctttccacc ccagctcaat ggcacagcca atgacacagc aggcccagag
     2941 ctgcctgacc ctttgggggg tctgttagat gaagctatgt tggatgagat cagccttatg
     3001 gacctggcca ttgaagaagg ctttaaccct gtgcaggcct cccagctgga ggaggaattt
     3061 gactctgact caggcctttc cttagactcg agccatagcc cttcttccct aagcagctct
     3121 gaaggcagtt cttcctcttc ttcctcctcc tcttcctctt cttcctctgc ttcttcctct
     3181 gcctcttcct ccttttctga ggaaggtgcg gttggctaca gctctgactc tgagaccctg
     3241 gatctggaag aggccgaggg tgctgtgggc taccagcctg agtattccaa gttctgccgc
     3301 atgagctacc aggatccagc tcagctctca tgcctgccct acctggagca cgtgggccac
     3361 aaccacacat acaacatggc acccagtgcc ctggactcag ccgacctgcc accacccagt
     3421 gccctcaaga aaggcagcaa ggagaagcag gctgacttcc tggacaagca gatgagccgg
     3481 gatgagcacc gagcccgagc catgaagatc cctttcacca atgacaaaat catcaacctg
     3541 cctgtggagg agttcaatga actgctgtcc aaataccagt tgagtgaagc ccagctgagc
     3601 ctcatccgag acatccggcg ccggggcaag aacaagatgg cggcgcagaa ctgccgcaag
     3661 cgcaagctgg acaccatcct gaatctggag cgtgatgtgg aggacctgca gcgtgacaaa
     3721 gcccggctgc tgcgggagaa agtggagttc ctgcgctccc tgcgacagat gaagcagaag
     3781 gtccagagcc tgtaccagga ggtgtttggg cggctgcgag atgagaacgg acgaccctac
     3841 tcgcccagtc agtatgcgct ccagtacgcc ggggacggca gtgtcctcct catcccccgc
     3901 acgatggccg accagcaggc ccggcggcag gagaggaagc caaaggaccg gagaaagtga
     3961 ggatcctgat atccagcaca gtggcggccg ctcgagtcta gagggcccgt ttaaacccgc
     4021 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     4081 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     4141 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     4201 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     4261 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     4321 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4381 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4441 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4501 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4561 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4621 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     4681 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     4741 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4801 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4861 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4921 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4981 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     5041 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     5101 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     5161 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     5221 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5281 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     5341 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5401 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5461 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5521 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5581 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5641 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     5701 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5761 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5821 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5881 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5941 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     6001 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     6061 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     6121 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     6181 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     6241 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6301 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6361 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6421 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6481 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6541 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6601 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6661 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6721 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6781 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6841 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6901 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6961 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7021 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7081 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     7141 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7201 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7261 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7321 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7381 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7441 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7501 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7561 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7621 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7681 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7741 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7801 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7861 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7921 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7981 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     8041 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8101 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8161 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8221 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8281 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8341 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8401 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     8461 ctatggt
//
