LOCUS       V026551                 8038 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026551
VERSION     V026551
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..3528
                     /label="SLC7A1(NM_003045)"
                     /note="SLC7A1(NM_003045)"
                     /gene="SLC7A1"
     polyA_signal    3605..3829
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3875..4303
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4317..4646
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4713..5507
                     /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    5681..5802
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5851..5867)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5875..5891
                     /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(5899..5929)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5944..5965
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6253..6838)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7009..7869)
                     /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(7870..7974)
                     /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 catggggtgc aaagtcctgc tcaacattgg gcagcagatg
     1681 ctgcggcgga aggtggtgga ctgtagccgg gaggagacgc ggctgtctcg ctgcctgaac
     1741 acttttgatc tggtggccct cggggtgggc agcacactgg gtgctggtgt ctacgtcctg
     1801 gctggagctg tggcccgtga gaatgcaggc cctgccattg tcatctcctt cctgatcgct
     1861 gcgctggcct cagtgctggc tggcctgtgc tatggcgagt ttggtgctcg ggtccccaag
     1921 acgggctcag cttacctcta cagctatgtc accgttggag agctctgggc cttcatcacc
     1981 ggctggaact taatcctctc ctacatcatc ggtacttcaa gcgtagcgag ggcctggagc
     2041 gccaccttcg acgagctgat aggcagaccc atcggggagt tctcacggac acacatgact
     2101 ctgaacgccc ccggcgtgct ggctgaaaac cccgacatat tcgcagtgat cataattctc
     2161 atcttgacag gacttttaac tcttggtgtg aaagagtcgg ccatggtcaa caaaatattc
     2221 acttgtatta acgtcctggt cctgggcttc ataatggtgt caggatttgt gaaaggatcg
     2281 gttaaaaact ggcagctcac ggaggaggat tttgggaaca catcaggccg tctctgtttg
     2341 aacaatgaca caaaagaagg gaagcccggt gttggtggat tcatgccctt cgggttctct
     2401 ggtgtcctgt cgggggcagc gacttgcttc tatgccttcg tgggctttga ctgcatcgcc
     2461 accacaggtg aagaggtgaa gaacccacag aaggccatcc ccgtggggat cgtggcgtcc
     2521 ctcttgatct gcttcatcgc ctactttggg gtgtcggctg ccctcacgct catgatgccc
     2581 tacttctgcc tggacaataa cagccccctg cccgacgcct ttaagcacgt gggctgggaa
     2641 ggtgccaagt acgcagtggc cgtgggctcc ctctgcgctc tttccgccag tcttctaggt
     2701 tccatgtttc ccatgcctcg ggttatctat gccatggctg aggatggact gctatttaaa
     2761 ttcttagcca acgtcaatga taggaccaaa acaccaataa tcgccacatt agcctcgggt
     2821 gccgttgctg ctgtgatggc cttcctcttt gacctgaagg acttggtgga cctcatgtcc
     2881 attggcactc tcctggctta ctcgttggtg gctgcctgtg tgttggtctt acggtaccag
     2941 ccagagcagc ctaacctggt ataccagatg gccagtactt ccgacgagtt agatccagca
     3001 gaccaaaatg aattggcaag caccaatgat tcccagctgg gctttttacc agaggcagag
     3061 atgttctctt tgaaaaccat actctcaccc aaaaacatgg agccttccaa aatctctggg
     3121 ctaattgtga acatttcaac cagcctcata gctgttctca tcatcacctt ctgcattgtg
     3181 accgtgcttg gaagggaggc tctcaccaaa ggggcgctgt gggcagtctt tctgctcgca
     3241 gggtctgccc tcctctgtgc cgtggtcacg ggcgtcatct ggaggcagcc cgagagcaag
     3301 accaagctct catttaaggt tcccttcctg ccagtgctcc ccatcctgag catcttcgtg
     3361 aacgtctatc tcatgatgca gctggaccag ggcacctggg tccggtttgc tgtgtggatg
     3421 ctgataggct tcatcatcta ctttggctat ggcctgtggc acagcgagga ggcgtccctg
     3481 gatgccgacc aagcaaggac tcctgacggc aacttggacc agtgcaagtg aggatcctga
     3541 tatccagcac agtggcggcc gctcgagtct agagggcccg tttaaacccg ctgatcagcc
     3601 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3661 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3721 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3781 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3841 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3901 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3961 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4021 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4081 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4141 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4201 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4261 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4321 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4381 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4441 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4501 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4561 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4621 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4681 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4741 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4801 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4861 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4921 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4981 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5041 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5101 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5161 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5221 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5281 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5341 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5401 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5461 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5521 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5581 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5641 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5701 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5761 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5821 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5881 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5941 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6001 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6061 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6121 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6181 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6241 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6301 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6361 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6421 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6481 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6541 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6601 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6661 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6721 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6781 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6841 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6901 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6961 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7021 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7081 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7141 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7201 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7261 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7321 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7381 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7441 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7501 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7561 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7621 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7681 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7741 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7801 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7861 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7921 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7981 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggt
//
