LOCUS       V039624                 8157 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039624
VERSION     V039624
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             884..1600
                     /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             1652..3646
                     /label="Slc5a1(NM_019810)"
                     /note="Slc5a1(NM_019810)"
                     /gene="Slc5a1"
     polyA_signal    3724..3948
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3994..4422
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4436..4765
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4832..5626
                     /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    5800..5921
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5970..5986)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5994..6010
                     /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(6018..6048)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6063..6084
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6372..6957)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7128..7988)
                     /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(7989..8093)
                     /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 ggctagcgct accggtcgcc accatggtga gcaagggcga
      901 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
      961 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     1021 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     1081 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1141 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     1201 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1261 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1321 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     1381 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     1441 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     1501 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1561 cgccgccggg atcactctcg gcatggacga gctgtacaag tccggactca gatctcgagc
     1621 cggtggaggc ggatcaggag gcggtggatc tatggacagt agcaccttga gccccgcggt
     1681 tactgccacc gatgcaccca tcccgtccta cgagcgcatt cgcaatgcgg ctgacatctc
     1741 agtcatcgtc atctacttcg tggtggtgat ggctgtcggg ctgtgggcca tgttttccac
     1801 taatcgtggt accgttggag gcttcttcct ggcaggccga agtatggtgt ggtggccgat
     1861 tggagcctct ctctttgcca gtaacattgg aagtggtcac tttgtggggc tggcaggcac
     1921 aggagcagcc gcaggcattg ccatgggtgg ctttgaatgg aacgccttgg ttttggtggt
     1981 tgtgctgggc tggatatttg tcccgattta catcaaggcc ggggtggtga cgatgccgga
     2041 gtatctgcgg aagcggtttg gaggcaagag aattcagatc tacctttccg ttctgtccct
     2101 gctcctctac atcttcacaa agatctctgc agacatcttc tccggggcta tattcatcaa
     2161 tctagccttg ggcctggata tatacttggc cattttcatc ctgctagcaa tcactgccct
     2221 ttatacaatc acaggggggc tggcagctgt aatttacaca gataccctgc agaccgcgat
     2281 catgctggtg gggtctttta tcctgactgg gtttgctttc aatgaagtag gaggatatga
     2341 agccttcatg gacaagtaca tgaaagctat tccaactaag gtttctaacg gaaattttac
     2401 cgccaaggaa gaatgctaca caccgagggc tgactcattc cacatattca gagaccccat
     2461 tactggagac atgccgtggc ctgggctcat ttttggcctg gccatcctcg ccctgtggta
     2521 ctggtgtacg gatcaggtca ttgtgcagcg atgcctctcg gccaagaaca tgtctcacgt
     2581 gaaggctggc tgtaccctgt gtggctatct gaagctactg cccatgttcc tcatggtgat
     2641 gccaggaatg atcagccgga ttctatatac agaaaaaatc gcctgtgtcc tccctgaaga
     2701 atgtcagaaa tactgcggca caccagtggg ctgtaccaac atcgcctacc ccacgctcgt
     2761 ggtggaactc atgcctaatg gactgcgagg cctgatgctg tctgtcatga tggcttccct
     2821 catgagctcc ttgacctcca tcttcaacag cgccagtact ctcttcacca tggacatcta
     2881 caccaagatc cggaagaagg catctgagaa ggagctcatg attgcaggaa ggttgttcat
     2941 cctcgtgctg attggcatca gcattgcctg ggtacccatt gtacagtctg ctcaaagtgg
     3001 gcagctcttt gattacatcc agtctatcac cagttacctg gggccgccca ttgcagctgt
     3061 cttcctgctt gccattttct gcaagagagt caatgaacaa ggagccttct ggggactgat
     3121 cctgggcttc ctgattggaa tctcccgtat gatcactgag tttgcctatg gaactgggag
     3181 ctgcatggag cccagcaact gccccaagat catctgtggg gtccactatt tgtactttgc
     3241 catcatcctc ttcgtcatca gcgtcatcac catcttgatc atctccttcc tcaccaagcc
     3301 cattccagac gtgcacctgt accgtttgtg ttggagtcta cgcaacagca aggaagagcg
     3361 aatcgacctg gatgcaggag aggaggagga catcccagag gactccaagg acacaattga
     3421 aatagacaca gaagcccccc agaagaagaa aggatgcttc aggagagcat atgacctgtt
     3481 ctgtgggctg gaccaggata aaggaccaaa gatgactaag gaggaggagg aagccatgaa
     3541 gatgaagatg acagacactt ctgagaagcc tttgtggagg acagtggtga acatcaacgg
     3601 tattatcctg ctggcggtag ctgtcttctg ccatgcctat tttgcctgag aattctgcag
     3661 tcgacggtac cgcgggcccg ggatcctcta gagggcccgt ttaaacccgc tgatcagcct
     3721 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     3781 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     3841 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     3901 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     3961 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     4021 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4081 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4141 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4201 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4261 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4321 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     4381 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     4441 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4501 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4561 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4621 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4681 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     4741 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     4801 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4861 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4921 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4981 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     5041 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     5101 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5161 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5221 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     5281 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5341 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5401 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5461 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     5521 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5581 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     5641 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     5701 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5761 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     5821 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5881 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     5941 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     6001 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     6061 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6121 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6181 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6241 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6301 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6361 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6421 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6481 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6541 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6601 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6661 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6721 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6781 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6841 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6901 ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6961 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7021 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7081 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7141 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7201 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7261 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7321 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7381 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7441 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7501 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7561 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7621 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7681 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7741 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7801 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7861 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7921 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7981 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     8041 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     8101 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggt
//
