LOCUS       V025091                 7049 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025091
VERSION     V025091
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..1729
                     /label="GALR1(NM_001480)"
                     /note="GALR1(NM_001480)"
                     /gene="GALR1"
     CDS             1736..1801
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1825..2413
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2479..2527
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2729..3157
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3171..3500
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3567..4361
                     /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    4537..4658
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4707..4723)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4731..4747
                     /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(4755..4785)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4800..4821
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5109..5697)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5868..6728)
                     /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(6729..6833)
                     /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 ccatggagct ggcggtcggg aacctcagcg agggcaacgc
      721 gagctggccg gagccccccg ccccggagcc cgggccgctg ttcggcatcg gcgtggagaa
      781 cttcgtcacg ctggtggtgt tcggcctgat cttcgcgctg ggtgtgctgg gcaacagcct
      841 agtgatcacc gtgctggcgc gcagcaagcc gggcaagccg cggagcacca ccaacctgtt
      901 catcctcaac ctgagcatcg ccgacctggc ctacctgctc ttctgcatcc ccttccaggc
      961 caccgtgtac gcgctgccca cctgggtgct gggcgccttc atctgcaagt tcatccacta
     1021 cttcttcacc gtgtccatgc tggtgagcat cttcaccctg gccgcgatgt ccgtggaccg
     1081 ctacgtggcc atcgtgcact cgcggcgctc ctcctccctc agggtgtccc gcaacgcgct
     1141 gctgggcgtg ggctgcatct gggcgctgtc cattgccatg gcctcgcccg tggcctacca
     1201 ccagggcctc ttccacccgc gcgccagcaa ccagaccttc tgctgggagc agtggcccga
     1261 ccctcgccac aagaaggcct acgtggtgtg caccttcgtc ttcggctacc tgctgccgct
     1321 cctgctcatc tgcttctgct atgccaaggt ccttaatcac ttgcataaaa agttgaagaa
     1381 catgtcaaag aagtctgaag catccaagaa aaagactgca cagacagttc tggtggtggt
     1441 tgtggtgttt ggaatctcct ggctgccgca ccacatcatc catctctggg ctgagtttgg
     1501 agttttcccg ctgacgccgg cttccttcct cttcagaatc accgcccact gcctggcgta
     1561 cagcaattcc tccgtgaatc ctatcattta tgcatttctc tctgaaaatt tcaggaaggc
     1621 ctataaacaa gtgttcaagt gtcacattcg caaagattca cacctgagtg atactaaaga
     1681 aagtaaaagt cgaatagaca ccccaccatc aaccaattgt actcatgtgc tcgaggacta
     1741 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa
     1801 gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     1861 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     1921 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     1981 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     2041 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2101 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2161 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
     2221 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     2281 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     2341 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     2401 gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg
     2461 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg
     2521 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     2581 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     2641 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg
     2701 cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2761 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2821 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2881 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2941 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3001 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3061 cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta
     3121 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3181 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3241 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3301 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3361 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3421 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3481 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3541 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3601 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3661 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3721 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3781 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3841 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3901 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3961 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4021 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4081 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4141 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4201 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4261 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4321 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc
     4381 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4441 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4501 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     4561 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4621 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     4681 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4741 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4801 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4861 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4921 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4981 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5041 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5101 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5161 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5221 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5281 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     5341 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5401 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5461 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5521 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     5581 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5641 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5701 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5761 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5821 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5881 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5941 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6001 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6061 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6121 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6181 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6241 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6301 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6361 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6421 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6481 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6541 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6601 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6661 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6721 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6781 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6841 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga
     6901 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     6961 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     7021 ccgacaattg catgaagaat ctgcttagg
//
