LOCUS       V044981                10066 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044981
VERSION     V044981
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..5556
                     /label="ABCC1(NM_004996)"
                     /note="ABCC1(NM_004996)"
                     /gene="ABCC1"
     CDS             5563..5628
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    5666..5890
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5936..6364
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6378..6707
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6774..7568
                     /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    7742..7863
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7912..7928)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7936..7952
                     /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(7960..7990)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8005..8026
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8314..8899)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9070..9930)
                     /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(9931..10035)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggcgc tccggggctt ctgcagcgcc gatggctccg acccgctctg ggactggaat
     1021 gtcacgtgga ataccagcaa ccccgacttc accaagtgct ttcagaacac ggtcctcgtg
     1081 tgggtgcctt gtttttacct ctgggcctgt ttccccttct acttcctcta tctctcccga
     1141 catgaccgag gctacattca gatgacacct ctcaacaaaa ccaaaactgc cttgggattt
     1201 ttgctgtgga tcgtctgctg ggcagacctc ttctactctt tctgggaaag aagtcggggc
     1261 atattcctgg ccccagtgtt tctggtcagc ccaactctct tgggcatcac catgctgctt
     1321 gctacctttt taattcagct ggagaggagg aagggagttc agtcttcagg gatcatgctc
     1381 actttctggc tggtagccct agtgtgtgcc ctagccatcc tgagatccaa aattatgaca
     1441 gccttaaaag aggatgccca ggtggacctg tttcgtgaca tcactttcta cgtctacttt
     1501 tccctcttac tcattcagct cgtcttgtcc tgtttctcag atcgctcacc cctgttctcg
     1561 gaaaccatcc acgaccctaa tccctgccca gagtccagcg cttccttcct gtcgaggatc
     1621 accttctggt ggatcacagg gttgattgtc cggggctacc gccagcccct ggagggcagt
     1681 gacctctggt ccttaaacaa ggaggacacg tcggaacaag tcgtgcctgt tttggtaaag
     1741 aactggaaga aggaatgcgc caagactagg aagcagccgg tgaaggttgt gtactcctcc
     1801 aaggatcctg cccagccgaa agagagttcc aaggtggatg cgaatgagga ggtggaggct
     1861 ttgatcgtca agtccccaca gaaggagtgg aacccctctc tgtttaaggt gttatacaag
     1921 acctttgggc cctacttcct catgagcttc ttcttcaagg ccatccacga cctgatgatg
     1981 ttttccgggc cgcagatctt aaagttgctc atcaagttcg tgaatgacac gaaggcccca
     2041 gactggcagg gctacttcta caccgtgctg ctgtttgtca ctgcctgcct gcagaccctc
     2101 gtgctgcacc agtacttcca catctgcttc gtcagtggca tgaggatcaa gaccgctgtc
     2161 attggggctg tctatcggaa ggccctggtg atcaccaatt cagccagaaa atcctccacg
     2221 gtcggggaga ttgtcaacct catgtctgtg gacgctcaga ggttcatgga cttggccacg
     2281 tacattaaca tgatctggtc agcccccctg caagtcatcc ttgctctcta cctcctgtgg
     2341 ctgaatctgg gcccttccgt cctggctgga gtggcggtga tggtcctcat ggtgcccgtc
     2401 aatgctgtga tggcgatgaa gaccaagacg tatcaggtgg cccacatgaa gagcaaagac
     2461 aatcggatca agctgatgaa cgaaattctc aatgggatca aagtgctaaa gctttatgcc
     2521 tgggagctgg cattcaagga caaggtgctg gccatcaggc aggaggagct gaaggtgctg
     2581 aagaagtctg cctacctgtc agccgtgggc accttcacct gggtctgcac gccctttctg
     2641 gtggccttgt gcacatttgc cgtctacgtg accattgacg agaacaacat cctggatgcc
     2701 cagacagcct tcgtgtcttt ggccttgttc aacatcctcc ggtttcccct gaacattctc
     2761 cccatggtca tcagcagcat cgtgcaggcg agtgtctccc tcaaacgcct gaggatcttt
     2821 ctctcccatg aggagctgga acctgacagc atcgagcgac ggcctgtcaa agacggcggg
     2881 ggcacgaaca gcatcaccgt gaggaatgcc acattcacct gggccaggag cgaccctccc
     2941 acactgaatg gcatcacctt ctccatcccc gaaggtgctt tggtggccgt ggtgggccag
     3001 gtgggctgcg gaaagtcgtc cctgctctca gccctcttgg ctgagatgga caaagtggag
     3061 gggcacgtgg ctatcaaggg ctccgtggcc tatgtgccac agcaggcctg gattcagaat
     3121 gattctctcc gagaaaacat cctttttgga tgtcagctgg aggaaccata ttacaggtcc
     3181 gtgatacagg cctgtgccct cctcccagac ctggaaatcc tgcccagtgg ggatcggaca
     3241 gagattggcg agaagggcgt gaacctgtct gggggccaga agcagcgcgt gagcctggcc
     3301 cgggccgtgt actccaacgc tgacatttac ctcttcgatg atcccctctc agcagtggat
     3361 gcccatgtgg gaaaacacat ctttgaaaat gtgattggcc ccaaggggat gctgaagaac
     3421 aagacgcgga tcttggtcac gcacagcatg agctacttgc cgcaggtgga cgtcatcatc
     3481 gtcatgagtg gcggcaagat ctctgagatg ggctcctacc aggagctgct ggctcgagac
     3541 ggcgccttcg ctgagttcct gcgtacctat gccagcacag agcaggagca ggatgcagag
     3601 gagaacgggg tcacgggcgt cagcggtcca gggaaggaag caaagcaaat ggagaatggc
     3661 atgctggtga cggacagtgc agggaagcaa ctgcagagac agctcagcag ctcctcctcc
     3721 tatagtgggg acatcagcag gcaccacaac agcaccgcag aactgcagaa agctgaggcc
     3781 aagaaggagg agacctggaa gctgatggag gctgacaagg cgcagacagg gcaggtcaag
     3841 ctttccgtgt actgggacta catgaaggcc atcggactct tcatctcctt cctcagcatc
     3901 ttccttttca tgtgtaacca tgtgtccgcg ctggcttcca actattggct cagcctctgg
     3961 actgatgacc ccatcgtcaa cgggactcag gagcacacga aagtccggct gagcgtctat
     4021 ggagccctgg gcatttcaca agggatcgcc gtgtttggct actccatggc cgtgtccatc
     4081 ggggggatct tggcttcccg ctgtctgcac gtggacctgc tgcacagcat cctgcggtca
     4141 cccatgagct tctttgagcg gacccccagt gggaacctgg tgaaccgctt ctccaaggag
     4201 ctggacacag tggactccat gatcccggag gtcatcaaga tgttcatggg ctccctgttc
     4261 aacgtcattg gtgcctgcat cgttatcctg ctggccacgc ccatcgccgc catcatcatc
     4321 ccgccccttg gcctcatcta cttcttcgtc cagaggttct acgtggcttc ctcccggcag
     4381 ctgaagcgcc tcgagtcggt cagccgctcc ccggtctatt cccatttcaa cgagaccttg
     4441 ctgggggtca gcgtcattcg agccttcgag gagcaggagc gcttcatcca ccagagtgac
     4501 ctgaaggtgg acgagaacca gaaggcctat taccccagca tcgtggccaa caggtggctg
     4561 gccgtgcggc tggagtgtgt gggcaactgc atcgttctgt ttgctgccct gtttgcggtg
     4621 atctccaggc acagcctcag tgctggcttg gtgggcctct cagtgtctta ctcattgcag
     4681 gtcaccacgt acttgaactg gctggttcgg atgtcatctg aaatggaaac caacatcgtg
     4741 gccgtggaga ggctcaagga gtattcagag actgagaagg aggcgccctg gcaaatccag
     4801 gagacagctc cgcccagcag ctggccccag gtgggccgag tggaattccg gaactactgc
     4861 ctgcgctacc gagaggacct ggacttcgtt ctcaggcaca tcaatgtcac gatcaatggg
     4921 ggagaaaagg tcggcatcgt ggggcggacg ggagctggga agtcgtccct gaccctgggc
     4981 ttatttcgga tcaacgagtc tgccgaagga gagatcatca tcgatggcat caacatcgcc
     5041 aagatcggcc tgcacgacct ccgcttcaag atcaccatca tcccccagga ccctgttttg
     5101 ttttcgggtt ccctccgaat gaacctggac ccattcagcc agtactcgga tgaagaagtc
     5161 tggacgtccc tggagctggc ccacctgaag gacttcgtgt cagcccttcc tgacaagcta
     5221 gaccatgaat gtgcagaagg cggggagaac ctcagtgtcg ggcagcgcca gcttgtgtgc
     5281 ctagcccggg ccctgctgag gaagacgaag atccttgtgt tggatgaggc cacggcagcc
     5341 gtggacctgg aaacggacga cctcatccag tccaccatcc ggacacagtt cgaggactgc
     5401 accgtcctca ccatcgccca ccggctcaac accatcatgg actacacaag ggtgatcgtc
     5461 ttggacaaag gagaaatcca ggagtacggc gccccatcgg acctcctgca gcagagaggt
     5521 cttttctaca gcatggccaa agacgccggc ttggtgctcg aggactacaa agaccatgac
     5581 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc
     5641 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     5701 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     5761 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     5821 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     5881 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     5941 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     6001 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     6061 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     6121 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     6181 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     6241 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     6301 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6361 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     6421 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     6481 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     6541 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     6601 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     6661 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     6721 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     6781 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     6841 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     6901 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     6961 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     7021 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     7081 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     7141 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     7201 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     7261 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     7321 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     7381 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     7441 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     7501 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     7561 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     7621 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     7681 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     7741 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     7801 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     7861 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     7921 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     7981 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     8041 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     8101 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     8161 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     8221 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     8281 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     8341 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     8401 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     8461 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     8521 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     8581 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     8641 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     8701 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     8761 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     8821 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     8881 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     8941 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     9001 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     9061 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     9121 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     9181 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     9241 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     9301 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     9361 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     9421 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     9481 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     9541 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     9601 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     9661 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     9721 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     9781 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     9841 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     9901 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     9961 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
    10021 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
