LOCUS       V029861                 7369 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029861
VERSION     V029861
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             706..2025
                     /label="GORASP1(NM_031899)"
                     /note="GORASP1(NM_031899)"
                     /gene="GORASP1"
     CDS             2032..2121
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2145..2733
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2799..2847
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3049..3477
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3491..3820
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3887..4681
                     /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    4857..4978
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5027..5043)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5051..5067
                     /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(5075..5105)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5120..5141
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5429..6017)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6188..7048)
                     /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(7049..7153)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatggg cctgggcgtc
      721 agcgctgagc agcccgcagg cggcgccgag ggcttccacc tccacggggt gcaggagaac
      781 tccccagccc agcaggcggg cctggagccc tactttgact tcatcatcac cattgggcac
      841 tcgaggctga acaaggagaa tgacaccctg aaggcactac tgaaagccaa tgtggagaag
      901 cccgtgaagc tggaggtgtt caatatgaag accatgaggg tgcgcgaggt ggaggtggtg
      961 cccagcaaca tgtggggcgg ccagggccta ctgggtgcca gtgtgcgctt ctgcagcttc
     1021 cgcagggcca gtgagcaggt gtggcatgtg ctggatgtgg aaccatcttc acctgctgcc
     1081 cttgccggcc tgcgccccta cacagactat gtggttggtt cggaccagat tctccaggag
     1141 tccgaggact tctttacgct catcgagtct catgagggga agcccttgaa gctgatggtg
     1201 tataactcca agtcagactc ctgccgggag gtgactgtaa ctcccaacgc agcctggggt
     1261 ggagagggca gtctgggatg tggcattggc tatgggtatc tacaccggat cccaactcag
     1321 ccccccagct accacaagaa gccacctggc accccaccac cttctgctct accacttggt
     1381 gccccaccac ctgatgctct accacctgga cccacccccg aggactctcc ttccctggag
     1441 acaggttcca ggcagagtga ctacatggag gccctgctgc aggcacctgg ctcctccatg
     1501 gaggatcccc ttcctgggcc tgggagtccc agccacagtg ctccagaccc tgatggactt
     1561 ccccatttca tggagactcc tcttcagccc ccacctccag tgcagcgagt tatggaccca
     1621 ggcttcctgg acgtgtcggg aatttctctc ttggacaaca gcaatgccag tgtgtggccc
     1681 agcctgccct cttccacaga actgaccacc acagctgtct caacctcagg gccagaggac
     1741 atctgctcca gcagcagttc tcatgagcgg ggtggtgagg ctacatggtc tgggtcagag
     1801 tttgaggtct ccttcctgga cagcccaggt gcccaagccc aggcggacca cctgcctcag
     1861 ctgactcttc ctgacagtct cacctctgca gcctcaccag aagatgggct gtccgccgag
     1921 ctgcttgaag ctcaggctga ggaggaacca gcaagcacag agggcctaga tactgggacg
     1981 gaggctgagg ggctggacag ccaggcccag atctctacca cagaactcga ggaacaaaaa
     2041 ctaatatcag aggaagattt ggaacagaaa ctaatcagtg aagaagattt agagcaaaag
     2101 ttaatttccg aggaggactt atagtgaggg cccgatatct cgacaatcaa cctctggatt
     2161 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     2221 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     2281 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     2341 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     2401 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     2461 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     2521 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
     2581 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     2641 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     2701 cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac
     2761 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     2821 aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     2881 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     2941 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     3001 gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta
     3061 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3121 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3181 ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc
     3241 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3301 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3361 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg
     3421 ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     3481 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3541 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3601 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3661 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3721 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     3781 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     3841 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3901 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3961 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4021 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4081 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4141 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4201 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4261 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4321 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4381 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4441 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4501 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4561 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4621 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4681 agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4741 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4801 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4861 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4921 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4981 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     5041 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5101 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5161 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5221 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5281 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5341 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5401 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5461 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5521 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5581 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt
     5641 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5701 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5761 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5821 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     5881 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5941 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6001 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6061 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6121 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6181 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6241 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6301 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6361 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6421 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6481 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6541 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6601 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6661 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6721 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6781 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6841 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6901 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6961 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7021 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7081 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7141 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     7201 tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca
     7261 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     7321 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
