LOCUS       V032009                 7862 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032009
VERSION     V032009
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..3352
                     /label="LRRC71(NM_144702)"
                     /note="LRRC71(NM_144702)"
                     /gene="LRRC71"
     CDS             3359..3424
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3462..3686
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3732..4160
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4174..4503
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4570..5364
                     /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    5538..5659
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5708..5724)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5732..5748
                     /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(5756..5786)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5801..5822
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6110..6695)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6866..7726)
                     /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(7727..7831)
                     /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 gagcgagcag agcgcgccgg gggcctcacc cagggccccg cgtccgggga cccagaagtc
     1741 ttctggcgcg gtgaccaaaa agggagagcg cgcggccaaa gagaagccag cgaccgttct
     1801 gcctcccgtg ggggaggagg agcccaaaag ccctgaggag taccagtgct ccggggtcct
     1861 cgagaccgac ttcgccgagc tctgcacgcg gtggggctac acggacttcc ccaaagttgt
     1921 caaccggccc cgcccccacc cgcccttcgt cccctccgcc tctttgtcgg aaaaggccac
     1981 cttagacgat ccgcggctgt cggggtcctg cagcctcaat agcctggaga gcaaatacgt
     2041 gttcttccgg cccaccatcc aggtggagct ggagcaggag gacagcaagt cagtgaagga
     2101 aatctacatc cgcggttgga aggttgagga acggattctg ggtgtcttct ctaaatgtct
     2161 gcccccgctt acccagctac aggccatcaa cttgtggaag gtggggctga ccgataagac
     2221 cctgaccacc ttcatcgagc tcctgcctct ctgttcatcc acgctcagga aggtgtctct
     2281 ggaggggaac ccactgccgg agcagtccta tcacaagctc atggccttgg acagcacgat
     2341 tgcgcacttg tctctgcgga acaataacat cgacgaccgc ggggcgcaac tcctgggcca
     2401 ggcgctgtcc acgctgcaca gctgcaaccg gaccctcgtc tcgctcaacc tgggtttcaa
     2461 ccacatcggt gacgagggcg caggctacat cgcggacggc ctccggctga accgttccct
     2521 gctctggctg tccctggccc acaaccgcat ccaggacaag ggcgccctga agctggctga
     2581 ggtcctgcgc gccttcgagc tgacacacac cgaagtggtg gagcgccgac gcctcctgct
     2641 ggaaaaaggg acacaggagc gctcgcgatc gccctcctcc tctcgacacg gggactccaa
     2701 aacggaccgt gagaagagtc agatggtagg gatcagcaat agtgcattgg tggacaagac
     2761 agacaagacg cagacaatga aaacccctaa gggcctgggc aagaaaaagg agaaatcatg
     2821 ggaattggcc aagaaagagg agaagttggg gtctgggcag tcacccacac aaggaacccc
     2881 taagaaggaa gatgccacaa aggcaggcaa ggggaaggta accatccctg aacagaagcc
     2941 aagcagggca aaagggatca agatcgggag cagagagaag cgcagcatcc tcctggagtc
     3001 cgagctggtt gttgaggcta ctgaggtggt caaccctctc ctggagcctg tggagcaccg
     3061 agatgggaaa gttttcatgc ctgggaacaa ggtccttttg cacctcaacc tcatccggaa
     3121 ccgcatcaca gaggtggggc tggagggctt cctcgccacg gtgcagtatc agatgcagtt
     3181 ctccaaggcc aagagtgcat ccaagggtcc agtggggctg ctgtggctgt ccctggctaa
     3241 aaattgcttc gccccacaat gtcctgcgta cgccataatc caggagctga tgttgccaag
     3301 ggatcccatc aaggccaaac tcagggagga tgaggccatg gcattcttcc ccctcgagga
     3361 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga
     3421 caagtagtga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     3481 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3541 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3601 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3661 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     3721 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3781 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3841 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3901 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3961 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4021 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4081 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4141 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4201 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4261 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4321 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4381 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4441 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4501 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4561 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4621 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4681 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4741 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4801 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4861 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4921 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4981 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5041 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5101 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5161 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5221 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5281 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5341 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5401 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5461 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5521 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5581 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5641 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5701 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5761 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5821 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5881 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5941 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6001 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6061 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6121 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6181 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6241 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6301 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6361 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6421 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6481 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6541 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6601 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     6661 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6721 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6781 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6841 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6901 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6961 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7021 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7081 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7141 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7201 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7261 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7321 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7381 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7441 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7501 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7561 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7621 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7681 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7741 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7801 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7861 tc
//
