LOCUS       V032652                 7859 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032652
VERSION     V032652
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             923..1639
                     /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             1676..3349
                     /label="Lrrc71(NM_028971)"
                     /note="Lrrc71(NM_028971)"
                     /gene="Lrrc71"
     CDS             3356..3421
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3459..3683
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3729..4157
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4171..4500
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4567..5361
                     /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    5535..5656
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5705..5721)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5729..5745
                     /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(5753..5783)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5798..5819
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6107..6692)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6863..7723)
                     /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(7724..7828)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
      961 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     1021 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     1081 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     1141 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     1201 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     1261 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     1321 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     1381 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     1441 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1501 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     1561 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     1621 catggacgag ctgtacaagg gcggtggagg cagtggtgga ggcggtagcg aattcatgtc
     1681 aagtgagcca agcactacag ggacctcacc caggactccg cgtcctgggg cccagaagtc
     1741 atccggagca gtaaccaaga agggggatcg ggctgctaaa gataagacag caagcaccct
     1801 gcccccggtg ggagaggacg agccaaaaaa tccggaggag taccagtgca cgggggtcct
     1861 ggagacagac ttcgctgagc tctgcacacg gtcaggctac gtggactttc ccaaagttgt
     1921 cacccggcca cgggtccaac agagctctgt cccttccgcc tccacgtcag aaaaacctgt
     1981 cctagatgac cagagaccgt cggcatcctg cagccagagc agcctggaga gcaagtatgt
     2041 gttttttcgg cccactattc aagtggagct ggagcaggag gacagcaagg ccgtgaagga
     2101 gatctacatt cgaggctgga aagttgaaga tcggattctg ggtattttct ccaaatgtct
     2161 gccctccctc agccagctgc aggccatcaa cttgtggaag gtggggctga cggataagac
     2221 cctgaccact ttcatcgccc ttttgcctct ttgttcctcc acactcagga aggtatctct
     2281 ggaggggaac ccaataccag agcagtcctt tagtaagctg atgggactgg acagcacgat
     2341 tgttcacttg tctctgagga acaacaacat caacgaccac ggggcgcagc tcctgggcca
     2401 ggcgctgtcc acactgcaga acagcaaccg aaccctggtt tcactgaacc tagcattcaa
     2461 ccacatcggg gatgtgggcg caggctacat tgcggatggc ttgaggctga accgctctct
     2521 cctctggctg tccctagcgc acaaccacat tcaagacaag ggagcactga agctggccga
     2581 ggtcctgcgc ccctttgagc tgacccacag ggaggtggtg gagcggcggc gcctgctgct
     2641 ggtgaaagga acgcaggagc ggtctcgatc gccttcctcc tcccgatacg gggactccaa
     2701 agcagaacgt gagaagtctc agacgttggg gatcagcaat gtcactttgg tggacaagca
     2761 agaaaagatg cagtcactga aagcccctaa gaccataagc aagaaaaagg agaagacagg
     2821 ggaagttgtc aagaaggaag agaagctggg ctctgggcag tcacccacac aaggaacccc
     2881 taagaaagaa gatgccgcaa aggcaggcaa ggggaaggta accatccctg agcagaagat
     2941 gagcaaggga aaagcgacaa agatgggggc caaagagaag cgcagcatcc tcctggagtc
     3001 agaacagctg gttgttgaag ctacagagat ggttaaccct ctcctggagc ccgtggaaca
     3061 ccgagatggg aaggtcttct tgcctgggaa caaggtcctt ttgcacctca acctcctccg
     3121 aaaccaaatc acagaggtgg ggctggaagg cttcctcact gctgtgcaat accaggttca
     3181 ggtctccaag cccaagactt cacccaaggc ccccctgggt ctgctgtggc tgtccttggc
     3241 gaaaaactgc tttgacccac agtgtccaac acacgtcatg attcaagaac tgatgttgcc
     3301 aagggaccct gtgaaggcca aggccaggga ggaggaggct gcagctaccc tcgaggacta
     3361 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa
     3421 gtagtgaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     3481 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3541 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3601 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3661 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3721 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3781 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3841 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3901 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3961 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4021 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4081 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4141 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4201 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4261 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4321 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4381 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4441 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4501 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4561 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4621 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4681 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4741 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4801 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4861 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4921 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4981 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5041 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     5101 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5161 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5221 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5281 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5341 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5401 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5461 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5521 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5581 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5641 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5701 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5761 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5821 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5881 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5941 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6001 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6061 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6121 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6181 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6241 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6301 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6361 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6421 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6481 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6541 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6601 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc
     6661 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6721 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6781 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6841 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6901 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6961 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7021 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7081 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7141 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7201 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7261 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7321 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7381 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7441 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7501 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7561 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7621 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7681 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7741 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7801 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
