LOCUS       V034765                 6286 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034765
VERSION     V034765
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..1776
                     /label="AQP2(NM_000486)"
                     /note="AQP2(NM_000486)"
                     /gene="AQP2"
     CDS             1783..1848
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    1886..2110
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2156..2584
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2598..2927
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             2994..3788
                     /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    3962..4083
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4132..4148)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4156..4172
                     /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(4180..4210)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4225..4246
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4534..5119)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5290..6150)
                     /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(6151..6255)
                     /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 accatgtggg agctccgctc catagccttc tccagggctg tgttcgcaga gttcctggcc
     1021 acactcctct tcgtcttctt tggcctcggc tctgccctca actggccaca ggccctgccc
     1081 tctgtgctac agattgccat ggcgtttggc ttgggtattg gcaccctggt acaggctctg
     1141 ggccacataa gcggggccca catcaaccct gccgtgactg tggcctgcct ggtgggctgc
     1201 cacgtctccg ttctccgagc cgccttctac gtggctgccc agctgctggg ggctgtggcc
     1261 ggagccgctc tgctccatga gatcacgcca gcagacatcc gcggggacct ggctgtcaat
     1321 gctctcagca acagcacgac ggctggccag gcggtgactg tggagctctt cctgacactg
     1381 cagctggtgc tctgcatctt cgcctccacc gatgagcgcc gcggagagaa cccgggcacc
     1441 cctgctctct ccataggctt ctctgtggcc ctgggccacc tccttgggat ccattacacc
     1501 ggctgctcta tgaatcctgc ccgctccctg gctccagctg tcgtcactgg caaatttgat
     1561 gaccactggg tcttctggat cggacccctg gtgggcgcca tcctgggctc cctcctctac
     1621 aactacgtgc tgtttccgcc agccaagagc ctgtcggagc gcctggcagt gctgaagggc
     1681 ctggagccgg acaccgattg ggaggagcgc gaggtgcgac ggcggcagtc ggtggagctg
     1741 cactcgccgc agagcctgcc acggggtacc aaggccctcg aggactacaa agaccatgac
     1801 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc
     1861 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     1921 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     1981 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2041 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     2101 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     2161 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2221 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2281 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2341 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     2401 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2461 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     2521 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2581 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2641 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     2701 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     2761 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2821 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     2881 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     2941 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     3001 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3061 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3121 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     3181 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3241 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3301 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     3361 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     3421 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     3481 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     3541 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     3601 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     3661 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     3721 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     3781 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     3841 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     3901 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     3961 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4021 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4081 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4141 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4201 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4261 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4321 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4381 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4441 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4501 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4561 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4621 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4681 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4741 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4801 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4861 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4921 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4981 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5041 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     5101 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5161 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5221 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5281 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5341 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5401 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5461 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5521 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5581 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5641 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5701 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5761 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5821 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5881 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5941 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6001 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6061 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6121 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6181 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6241 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
