LOCUS       V040230                 7788 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040230
VERSION     V040230
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2468
                     /label="Apba3(NM_018758)"
                     /note="Apba3(NM_018758)"
                     /gene="Apba3"
     CDS             2475..2540
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2564..3152
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3218..3266
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3468..3896
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3910..4239
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4306..5100
                     /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    5276..5397
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5446..5462)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5470..5486
                     /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(5494..5524)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5539..5560
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5848..6436)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6607..7467)
                     /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(7468..7572)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgga gttcctgcca ggaccccagc
      781 atcccccagg gcccccaacc atggacttgg aggagcccaa aggccccgag gtgccctctg
      841 agaaccaccc ctctaacacc cagtcggcct tagggcctgg aggccctgtc accctgagtg
      901 agatggagct ggacacatcc agtgtgcagg agctggttca gcagctagaa gctctaccca
      961 gcgacctggg tggcccattc ccagatggag ccccctgtcc cctgcacatt gctaccggcc
     1021 aaggacttgc cacccaggag aacccagatg ctggtgggct cttgtcggcc gaggcaggcg
     1081 gggacgacct tctgggtctg ctgagggatg aggcctcttc gcctgcccaa tctgtccccc
     1141 aagatcctgc acagactgca ccccgccttc tgcagccgcc agaggaccct gatggggacc
     1201 cgggatggat ggaaggggca tcagcagagc cagcagacag caggagctcc agcagttccc
     1261 cggagccctg gctggagaca gcacccctgg tgacacagca ggaaccacct gtgggtaccc
     1321 agagccggga gaccctggcc tcgtgccctg ccgtcaccga ggtcccaggt ccctgtgggc
     1381 cggaggagct catggatggt gtcatcttcg gggcgaaata cctgggctcc acccagctgc
     1441 tgtcagagcg cagcccggcg cccagcacac gcatggggca ggcgcaggag gccatggacc
     1501 gcgtcaaggc ccctgaaggc gagacccagc ccatggtgga agtggacatc tttatctcca
     1561 ccaagcgtgt caaagtgttg gctgctgact cccaggatgc cctgatggac catgccctgc
     1621 agaccatctc ctacatcgca gacattgggc ctgtactggt cttgatggcc cggaggcggc
     1681 tggccaggag gacaactccc caggaccgcc agcggcgact ctacaagatg ctgtgccatg
     1741 tcttccactc agaggatgcc cagctcatag cacaagccat cggccaggcc ttcagcatcg
     1801 cctacagcca gttcctacag gagaacagga tcgaccccag ccaggtgggc acgcagccct
     1861 caaccgctgc cagccacccg cacaacggcg acctggacca cttctgcaac agccagaact
     1921 gtagggaggt ttgcatccag aagcggccag gtgaaggcct gggcgtcgcg ctagttgaat
     1981 caggctgggg ctcgctactg cccactgcgg tcattgccaa cctgctccat gggggccctg
     2041 ccgagcgctg cggggccctg agcatagggg accgcgtcac tgccatcaat ggaaccagcc
     2101 tggtggggct gtcgctggct gcctgccagg ctgcggtgcg cgaggtgcgg cgactttcat
     2161 ccgtgacgct gagtattatc cactgcccac cggtcaccac ggccgtcatc cgccggcccc
     2221 acgtgcgcga gcagctgggc ttctgtgtgg aggatggaat catttgtagc ctgctgcgtg
     2281 ggggcgctgc tgagcgtggt ggagtccgcg ttggacaccg catcattgag gtcaacgggc
     2341 agagcgtggt ggctatgccc catgcgcgca tcatccagct gctcacagag acaagagaaa
     2401 tccacatcaa gaccatgcca gctgccacct accgactgct cacgggacag gaacaaccag
     2461 tatacctagg atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact
     2521 acaaggatga cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta
     2581 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2641 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2701 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     2761 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2821 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     2881 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     2941 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     3001 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     3061 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3121 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca
     3181 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     3241 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     3301 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     3361 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     3421 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     3481 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3541 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3601 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     3661 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3721 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3781 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     3841 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3901 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3961 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4021 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4081 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4141 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4201 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4261 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4321 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4381 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4441 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4501 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4561 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4621 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4681 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4741 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4801 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4861 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4921 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4981 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5041 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5101 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5161 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5221 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5281 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5341 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5401 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5461 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5521 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5581 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5641 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5701 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5761 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5821 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5881 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5941 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6001 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     6061 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6121 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6181 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6241 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6301 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6361 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6421 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6481 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6541 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6601 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6661 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6721 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6781 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6841 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6901 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6961 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7021 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7081 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7141 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7201 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7261 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7321 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7381 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7441 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7501 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7561 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     7621 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag
     7681 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     7741 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
