LOCUS       V020588                 7271 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020588
VERSION     V020588
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..1951
                     /label="GHRHR(NM_000823)"
                     /note="GHRHR(NM_000823)"
                     /gene="GHRHR"
     CDS             1958..2023
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2047..2635
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2701..2749
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2951..3379
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3393..3722
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3789..4583
                     /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    4759..4880
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4929..4945)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4953..4969
                     /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(4977..5007)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5022..5043
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5331..5919)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6090..6950)
                     /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(6951..7055)
                     /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 ccatggaccg ccggatgtgg ggggcccacg tcttctgcgt
      721 gttgagcccg ttaccgaccg tattgggcca catgcaccca gaatgtgact tcatcaccca
      781 gctgagagag gatgagagtg cctgtctaca agcagcagag gagatgccca acaccaccct
      841 gggctgccct gcgacctggg atgggctgct gtgctggcca acggcaggct ctggcgagtg
      901 ggtcaccctc ccctgcccgg atttcttctc tcacttcagc tcagagtcag gggctgtgaa
      961 acgggattgt actatcactg gctggtctga gccctttcca ccttaccctg tggcctgccc
     1021 tgtgcctctg gagctgctgg ctgaggagga atcttacttc tccacagtga agattatcta
     1081 caccgtgggc catagcatct ctattgtagc cctcttcgtg gccatcacca tcctggttgc
     1141 tctcaggagg ctccactgcc cccggaacta cgtccacacc cagctgttca ccacttttat
     1201 cctcaaggcg ggagctgtgt tcctgaagga tgctgccctt ttccacagcg acgacactga
     1261 ccactgcagc ttctccactg ttctatgcaa ggtctctgtg gccgcctccc atttcgccac
     1321 catgaccaac ttcagctggc tgttggcaga agccgtctac ctgaactgcc tcctggcctc
     1381 cacctccccc agctcaagga gagccttctg gtggctggtt ctcgctggct gggggctgcc
     1441 cgtgctcttc actggcacgt gggtgagctg caaactggcc ttcgaggaca tcgcgtgctg
     1501 ggacctggac gacacctccc cctactggtg gatcatcaaa gggcccattg tcctctcggt
     1561 cggggtgaac tttgggcttt ttctcaatat tatccgcatc ctggtgagga aactggagcc
     1621 agctcagggc agcctccata cccagtctca gtattggcgt ctctccaagt cgacactttt
     1681 cctgatccca ctctttggaa ttcactacat catcttcaac ttcctgccag acaatgctgg
     1741 cctgggcatc cgcctccccc tggagctggg actgggttcc ttccagggct tcattgttgc
     1801 catcctctac tgcttcctca accaagaggt gaggactgag atctcacgga agtggcatgg
     1861 ccatgaccct gagcttctgc cagcctggag gacccgtgct aagtggacca cgccttcccg
     1921 ctcggcggca aaggtgctga catctatgtg cctcgaggac tacaaagacc atgacggtga
     1981 ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgatat
     2041 ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     2101 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     2161 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     2221 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     2281 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     2341 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     2401 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     2461 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     2521 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     2581 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga
     2641 aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     2701 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg
     2761 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     2821 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     2881 gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg
     2941 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3001 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3061 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg
     3121 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3181 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3241 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3301 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa
     3361 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3421 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3481 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3541 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3601 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3661 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3721 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     3781 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3841 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3901 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3961 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4021 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4081 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4141 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4201 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4261 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4321 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4381 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4441 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4501 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4561 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg
     4621 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4681 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4741 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4801 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4861 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4921 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4981 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5041 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5101 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5161 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5221 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5281 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5341 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5401 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5461 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5521 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5581 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5641 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5701 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5761 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5821 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5881 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5941 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6001 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6061 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6121 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6181 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6241 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6301 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6361 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6421 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6481 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6541 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6601 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6661 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6721 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6781 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6841 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6901 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6961 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7021 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7081 gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct
     7141 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     7201 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     7261 atctgcttag g
//
