LOCUS       V020773                 6998 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020773
VERSION     V020773
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             683..1678
                     /label="Mc4r(NM_013099)"
                     /note="Mc4r(NM_013099)"
                     /gene="Mc4r"
     CDS             1685..1750
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1774..2362
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2428..2476
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2678..3106
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3120..3449
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3516..4310
                     /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    4486..4607
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4656..4672)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4680..4696
                     /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(4704..4734)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4749..4770
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5058..5646)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5817..6677)
                     /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(6678..6782)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgaactc cacccaccac catggcatgt atacttccct
      721 ccacctctgg aaccgcagca gccacgggct gcacggcaat gccagcgagt ctctggggaa
      781 ggggcactca gacggaggat gctatgagca actttttgtc tcccccgagg tgtttgtgac
      841 tctgggtgtc ataagcctgt tggagaacat tctagtgatc gtggcgatag ccaagaacaa
      901 gaacctgcac tcacccatgt actttttcat ctgtagtctg gctgtggcgg acatgctggt
      961 gagcgtttcg aacgggtcag aaaccatcgt catcaccctg ctaaacagta cggacacgga
     1021 cgcccagagc ttcaccgtga atattgataa tgtcattgac tctgtgatct gtagctcctt
     1081 gctcgcatcc atttgcagcc tgctttccat tgcagtggac aggtatttca ctatctttta
     1141 cgcgctccag taccataaca ttatgacggt taggcgggtc gggatcatca tcagttgtat
     1201 ctgggcagct tgcacagtat cgggcgttct ttttatcatt tactcggaca gcagcgctgt
     1261 catcatctgc ctcattacca tgttcttcac catgctggtt ctcatggcct ctctctatgt
     1321 ccacatgttc ctgatggcga ggcttcacat taagaggatc gctgtcctcc cgggcacggg
     1381 taccatccgt cagggtgcca acatgaaggg cgcaattacc ttgaccattc tgattggagt
     1441 gtttgttgtc tgctgggccc cgtttttcct ccatttactg ttctacatct cttgtcctca
     1501 gaatccatac tgcgtgtgct tcatgtctca ttttaacttg tatctcatac tgatcatgtg
     1561 taacgctgtc atcgaccctc tcatttatgc cctgcggagt caagaactga ggaaaacctt
     1621 caaagagatc atctgtttct accccctggg aggcatctgt gagttacctg gcaggtatct
     1681 cgaggactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     1741 cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta caaaatttgt
     1801 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     1861 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     1921 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     1981 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     2041 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     2101 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     2161 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     2221 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     2281 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     2341 tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga
     2401 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt
     2461 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     2521 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     2581 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     2641 gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     2701 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2761 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2821 gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2881 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2941 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3001 acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc
     3061 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3121 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3181 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3241 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3301 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3361 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3421 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3481 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3541 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3601 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3661 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3721 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3781 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3841 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3901 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3961 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4021 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     4081 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4141 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4201 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4261 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4321 ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4381 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4441 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4501 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4561 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4621 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4681 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4741 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4801 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4861 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4921 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4981 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5041 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5101 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5161 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5221 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     5281 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5341 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5401 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5461 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     5521 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5581 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5641 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5701 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5761 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5821 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5881 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5941 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6001 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6061 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6121 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6181 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6241 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6301 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6361 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6421 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6481 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6541 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6601 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6661 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6721 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6781 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     6841 tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     6901 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     6961 aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
