LOCUS       V031446                 8024 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031446
VERSION     V031446
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             929..3514
                     /label="RABEP1(NM_004703)"
                     /note="RABEP1(NM_004703)"
                     /gene="RABEP1"
     CDS             3521..3586
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3624..3848
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3894..4322
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4336..4665
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4732..5526
                     /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    5700..5821
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5870..5886)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5894..5910
                     /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(5918..5948)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5963..5984
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6272..6857)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7028..7888)
                     /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(7889..7993)
                     /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 aagcttggta ccgccaccat ggcgcagccg ggcccggctt cccagcctga
      961 cgtttctctt cagcaacggg tagcagaatt ggaaaaaatt aatgcagaat ttttacgtgc
     1021 acaacagcag cttgaacaag aatttaatca aaagagagca aaatttaagg agttatattt
     1081 ggctaaagag gaggatctga agaggcaaaa tgcagtatta caagctgcac aagatgattt
     1141 gggacacctt cgaacccagc tgtgggaagc tcaagcagag atggagaata ttaaggcgat
     1201 tgccacagtc tctgagaaca ccaagcaaga agctatagat gaagtgaaaa gacagtggag
     1261 agaagaagtt gcttcacttc aggctgttat gaaagaaaca gttcgtgact atgagcacca
     1321 gttccacctt aggctggagc aggagcgaac acagtgggca cagtatagag aatccgcaga
     1381 gagggaaata gctgatttaa gaagaaggct gtctgaaggt caagaggagg aaaatttaga
     1441 aaatgaaatg aaaaaggccc aagaggatgc tgagaaactt cggtccgttg tgatgccaat
     1501 ggaaaaggaa attgcagctt tgaaggataa actgacagag gctgaagaca aaattaaaga
     1561 gctggaggcc tcaaaggtta aagaactgaa tcattatctg gaagctgaga aatcttgtag
     1621 gactgatcta gagatgtatg tagctgtttt gaatactcag aaatctgttc tacaggaaga
     1681 tgctgagaaa ctgcggaaag aattgcatga agtttgccat ctcttggagc aagagcgaca
     1741 acaacacaac cagttaaaac atacgtggca gaaggccaat gaccagtttc tggaatctca
     1801 gcgtttactg atgagagaca tgcagcgaat ggagattgtg ctaacttcag aacagctccg
     1861 acaagttgaa gaactgaaga agaaagatca ggaggatgat gaacaacaaa gactcaataa
     1921 gagaaaggat cacaaaaaag cagatgttga ggaagaaata aaaataccag tagtgtgtgc
     1981 tttaactcaa gaagaatctt cagcccagtt atcaaatgaa gaggagcatt tagacagcac
     2041 ccgtggctca gttcattcct tagatgcagg cttgctgttg ccatctggag atcctttcag
     2101 taaatcggac aatgacatgt ttaaagatgg actcaggaga gcacagtcta cagacagctt
     2161 gggaacctcg ggctcattgc aatccaaagc tttaggctat aactacaaag caaaatctgc
     2221 tggaaacctg gacgagtcag attttggacc actggtagga gcagattcag tgtctgagaa
     2281 ctttgatact gcatcccttg ggtcactcca gatgccaagt gggtttatgt taaccaaaga
     2341 tcaggaaaga gcaatcaagg cgatgacacc agaacaagaa gagacagcgt ccctcctctc
     2401 cagcgttacc cagggcatgg agagtgccta tgtgtcccct agtggttatc gtttagttag
     2461 tgaaacagaa tggaatctct tgcagaaaga ggtacataat gctggaaata aacttggtag
     2521 acgttgtgat atgtgttcca attacgaaaa acagttacaa ggaattcaga ttcaggaggc
     2581 tgaaacgaga gaccaggtga aaaaactaca gctgatgcta aggcaagcta atgaccagtt
     2641 agagaagaca atgaaagata agcaggagct ggaagacttc ataaagcaaa gcagcgaaga
     2701 ttcgagtcac cagatctctg cactcgtcct aagagcccag gcctccgaga tcttacttga
     2761 agagttacag caggggcttt cccaggcaaa gagggatgtt caggaacaga tggcggtgct
     2821 gatgcagtca cgggaacagg tttcagaaga gctggtgagg ttacagaaag ataatgacag
     2881 tctccaggga aagcacagcc tgcatgtgtc attacagcaa gcagaagact tcatcctccc
     2941 agacactaca gaggcactgc gggagttggt attaaaatac cgcgaggaca tcattaatgt
     3001 gcggacagca gcagaccacg tagaagaaaa gctgaaggct gagatacttt tcctaaaaga
     3061 gcagatccaa gcagaacagt gtttaaaaga aaatcttgaa gaaactctgc aactagaaat
     3121 agaaaactgc aaggaggaaa tagcttctat ttctagccta aaagctgaat tagaaagaat
     3181 aaaagtggaa aaaggacagt tggagtccac attaagagag aagtctcaac agcttgagag
     3241 tcttcaggaa ataaagatca gtttggaaga gcagttaaag aaagagactg ctgctaaggc
     3301 taccgttgaa cagctaatgt ttgaagagaa gaacaaagct cagagattac agacagaatt
     3361 agatgtcagt gagcaagtcc agagagattt tgtaaagctt tcacagaccc ttcaggtgca
     3421 gttagagcgg atccggcaag ctgactcctt ggagagaatc cgggcaattc tgaatgatac
     3481 taaactgaca gacattaacc agcttcctga gacactcgag gactacaaag accatgacgg
     3541 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt
     3601 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3661 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3721 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3781 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3841 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3901 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3961 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4021 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4081 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4141 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4201 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4261 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4321 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4381 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4441 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4501 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4561 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4621 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4681 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4741 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4801 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4861 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4921 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4981 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5041 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5101 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5161 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5221 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     5281 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5341 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5401 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5461 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5521 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5581 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5641 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5701 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5761 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5821 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5881 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5941 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6001 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6061 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6121 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6181 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6241 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6301 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6361 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6421 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6481 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6541 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6601 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6661 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6721 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6781 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca
     6841 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6901 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6961 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7021 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7081 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7141 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7201 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7261 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7321 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7381 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7441 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7501 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7561 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7621 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7681 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7741 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7801 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7861 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7921 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7981 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
