LOCUS       V025176                 7582 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025176
VERSION     V025176
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..2241
                     /label="Slc7a5(NM_011404)"
                     /note="Slc7a5(NM_011404)"
                     /gene="Slc7a5"
     CDS             2248..2274
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2278..2304
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2308..2334
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2358..2946
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3012..3060
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3262..3690
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3704..4033
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4100..4894
                     /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    5070..5191
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5240..5256)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5264..5280
                     /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(5288..5318)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5333..5354
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5642..6230)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6401..7261)
                     /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(7262..7366)
                     /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 ccaccatggc ggtcgcgggc
      721 gccaagaggc gcgcggttgc gaccccggcg gcggcggcgg cggaggagga gcggcaggcg
      781 cgggagaaga tgttggaggc gcggcgcggg gacggcgcgg accccgaggg cgaaggcgtg
      841 accctgcagc gcaatatcac gctgctcaac ggtgtggcca tcatcgtggg caccatcatc
      901 ggctctggca tcttcgtgac gcccaccggc gtgctcaagg aagcggggtc gcccggactg
      961 tcgctcgtgg tgtgggctgt gtgcggcgtc ttctccatcg tgggcgcact ctgctacgcg
     1021 gagctgggca ccaccatctc caagtcaggc ggcgactacg cctacatgct ggaggtctac
     1081 ggctcgttgc ccgccttcct caagctctgg atcgagctgc tcatcattcg gccctcctcg
     1141 cagtacatcg tggcgctggt cttcgccacc tacttgctca agccggtctt ccccacttgt
     1201 ccggtgcccg aggaagctgc caagctcgtg gcctgcctct gcgtgctgct cctcacagct
     1261 gtgaactgct acagcgtaaa ggctgcgacc cgtgtgcagg acgcctttgc ggctgccaag
     1321 ctgctggccc tggccctcat cattttgctc ggcttcatcc agatggggaa ggacatggga
     1381 caaggtgatg cgtccaacct gcagcagaag ttgtcctttg aaggcaccaa tctggacgtg
     1441 gggaacattg tgttggcttt gtacagcggc ctcttcgcct acggaggatg gaactatctg
     1501 aattttgtca cggaggagat gatcaacccc tacaggaacc tgcccctggc catcatcatc
     1561 tccttgccca ttgtcacact ggtctatgtg ctgacgaacc tggcctattt cactaccctc
     1621 tctaccaacc agatgctgac atctgaagct gtggctgtgg acttcgggaa ctaccacttg
     1681 ggtgtcatgt cctggatcat tcctgtcttt gtgggcttgt cttgcttcgg ctctgtcaat
     1741 gggtctctgt tcacgtcctc aagactgttc ttcgtgggat ccagggaggg ccacctgcct
     1801 tctgtcctct ccatgatcca cccacagctg ctgacacctg tgccatcact ggtgttcacg
     1861 tgtatcatga cgctgatgta cgccttctcc agagatatct tctccatcat caacttcttc
     1921 agcttcttca actggctgtg tgtggccctg gccatcatcg gtatgatgtg gctccgattc
     1981 aagaagcctg agctggagcg tcccatcaag gtgaatctgg ccctcccggt gttctttatc
     2041 ctggcctgcc tcttcctcat tgccgtgtca ttctggaaga cacccatgga gtgtggcatt
     2101 ggcttcgcca tcatcctcag cgggctgcct gtctacttct ttggtgtctg gtggaagaac
     2161 aagcccaagt ggattctgca ggctatcttc tctgtgacgg tgctctgcca gaagttgatg
     2221 caggtggtac ctcaggagac tctcgagtac ccatacgacg tacctgatta tgcaacttat
     2281 ccttatgatg tacctgatta tgctagctat ccttatgatg tcccagacta cgcatagtga
     2341 gggcccgata tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     2401 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     2461 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     2521 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     2581 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     2641 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     2701 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     2761 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     2821 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     2881 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     2941 ccccgcctgg aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     3001 ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc
     3061 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     3121 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     3181 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct
     3241 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3301 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3361 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca
     3421 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3481 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3541 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3601 cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg
     3661 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     3721 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3781 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3841 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3901 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3961 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4021 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     4081 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4141 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4201 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4261 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4321 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4381 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4441 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4501 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4561 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4621 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4681 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4741 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4801 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4861 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga
     4921 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4981 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5041 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     5101 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5161 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     5221 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5281 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5341 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5401 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5461 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5521 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5581 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5641 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5701 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5761 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5821 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5881 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5941 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6001 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6061 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     6121 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6181 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6241 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6301 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6361 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6421 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6481 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6541 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6601 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6661 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6721 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6781 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6841 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6901 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6961 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7021 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7081 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7141 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7201 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7261 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7321 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7381 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag
     7441 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     7501 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     7561 ttgcatgaag aatctgctta gg
//
