LOCUS       V035832                 9038 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035832
VERSION     V035832
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             884..1591
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             1628..4558
                     /label="INPP4A(NM_001134224)"
                     /note="INPP4A(NM_001134224)"
                     /gene="INPP4A"
     polyA_signal    4605..4829
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4875..5303
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5317..5646
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5713..6507
                     /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    6681..6802
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6851..6867)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6875..6891
                     /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(6899..6929)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6944..6965
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7253..7838)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8009..8869)
                     /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(8870..8974)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgct accggtcgcc accatggtga gcaagggcga
      901 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
      961 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     1021 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     1081 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     1141 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     1201 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     1261 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     1321 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     1381 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     1441 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     1501 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     1561 ctccaccggc ggcatggacg agctgtacaa gggtggaggc ggatcaggag gcggtggatc
     1621 tctcgagatg acagcaagag agcacagccc tcgccatggt gccagggccc gtgcaatgca
     1681 gcgggcttcc accatcgacg tggcggccga catgctgggc ctctctctgg caggaaatat
     1741 acaagaccca gatgagccca ttttagaatt tagcttagct tgcagtgagc tgcatactcc
     1801 atcgctagat cgaaagccaa atagttttgt tgcggtgagt gtcaccaccc ctcctcaggc
     1861 attctggacg aagcatgcac agacggagat cattgaggga accaacaatc ctatatttct
     1921 aagcagtatt gccttctttc aagactctct tatcaatcag atgacacaag tcaaactctc
     1981 cgtgtatgat gtcaaagata gatctcaggg aacaatgtat ttactgggct ctggaacgtt
     2041 cattgtcaaa gatctgctcc aggacaggca tcataggttg catttaacac taaggtctgc
     2101 agagagtgac cgtgtaggta acatcaccgt gattggctgg cagatggagg agaagtcaga
     2161 ccaacggccc cctgtgaccc ggtctgtgga cactgtcaat gggaggatgg ttcttcctgt
     2221 cgatgagagc ttgacggagg cgttaggaat ccgatccaaa tacgcttcat tgcgaaagga
     2281 cactttgctg aaatcggtgt tcggtggtgc catctgccgc atgtaccggt ttccaaccac
     2341 tgatggtaac catttgcgga tcctggagca gatggcagag agcgtgctct ccctgcacgt
     2401 gccccggcag ttcgtgaagc tcctactaga ggaagatgca gccagagtgt gtgagctgga
     2461 ggagctggga gagctgtccc cttgctggga gagcctccgg cgccaaattg tcacccagta
     2521 ccagaccatc atcctcacat accaggagaa cctgaccgac ctccatcagt acagagggcc
     2581 ctcgtttaaa gcaagcagtt tgaaagcaga taaaaagtta gaatttgttc ccacaaactt
     2641 gcacatacaa aggatgagag ttcaagacga tggaggatca gatcagaact acgacatcgt
     2701 caccattggg gcgccagcag cacactgcca aggttttaag tcaggaggtc tccgcaaaaa
     2761 gctgcacaaa tttgaagaga ccaagaaaca ttttgaggag tgttgtacat catctggctg
     2821 ccagtccata atctacatac cccaggatgt tgtcagagcc aaggagatca tcgcccagat
     2881 caacaccctg aaaacccaag tgagttacta cgcagagcgg ctgtcaaggg cagccaagga
     2941 caggtctgcc actggccttg agaggacact cgccatcttg gcagacaaga cacggcagct
     3001 ggtcacggtc tgcgactgca agctcctggc caactccatc catgggctga acgctgcacg
     3061 gcctgactac attgcctcca aggcctctcc cacttcgact gaggaggagc aggtgatgct
     3121 tagaaatgac caggacaccc tcatggcccg gtggacaggg agaaacagcc gatcttccct
     3181 gcaggtggac tggcacgagg aggagtggga gaaagtgtgg ctgaacgtgg acaagagcct
     3241 agagtgcatc attcagcgtg tggacaagct gctgcagaag gagcggctgc atggcgaggg
     3301 ctgtgaggat gtcttcccct gtgcaggcag ctgcaccagc aagaaaggta acccggacag
     3361 ccacgcctac tggatcagac cagaagaccc cttctgtgat gtcccctcct caccatgccc
     3421 ctccaccatg ccctccactg catgccatcc tcatctgacc acacattgca gtccccctcc
     3481 tgaagagtcc agcccaggtg aatggagtga ggccctttac ccgctgctga ccactctcac
     3541 cgactgcgtg gccatgatga gtgacaaggc caagaaggcc atggtattcc tgctcatgca
     3601 ggacagcgcg cccaccatag ccacctacct gagcctgcag taccgccgtg acgtggtctt
     3661 ctgccagacg ctgaccgccc tcatctgcgg cttcatcatt aagctgagga actgcctgca
     3721 tgacgacggc ttcctgcgcc agctctacac catcgggctg ctggcccagt tcgagagcct
     3781 gctgagcacc tacggggagg agctggcaat gctggaggac atgagccttg ggatcatgga
     3841 cttgaggaac gtgaccttca aagtcactca ggccacttcc agcgcctccg cagacatgct
     3901 gcccgtcatc acaggaaatc gcgacgggtt taacgtgcgg gtccctctgc cgggcccgct
     3961 gtttgacgcc ttgccccggg agatccagag tggcatgctg ctgcgagtgc agcccgtcct
     4021 cttcaacgtg ggcatcaatg agcagcagac actggccgag aggtttggcg atacgtcttt
     4081 acaagaagtc atcaacgtgg agagtttggt gcggttaaat tcctactttg agcagtttaa
     4141 ggaagttttg cctgaggatt gcctgcctcg gtctcgcagt cagacgtgcc tgccagagct
     4201 gctgcggttt ctgggtcaga acgtgcatgc ccggaagaat aagaacgtcg acattctctg
     4261 gcaagctgct gagatctgcc gccgccttaa tggggtccgg ttcaccagct gcaagagcgc
     4321 taaggaccgt acagccatgt cggtgacact ggagcagtgc ctgatcctgc aacacgagca
     4381 tggcatggcc ccgcaggtct tcacccaggc cctggagtgc atgcgcagtg agggttgtcg
     4441 aagagaaaat acaatgaaga atgttggaag tcgcaaatat gcatttaatt ccctgcagct
     4501 gaaggctttc cccaagcatt acaggcctcc cgaagggact tacggaaaag ttgaaacgtg
     4561 aggatcctct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     4621 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     4681 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     4741 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     4801 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     4861 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4921 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4981 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     5041 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     5101 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     5161 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     5221 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     5281 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     5341 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5401 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5461 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     5521 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     5581 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     5641 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     5701 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5761 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5821 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5881 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5941 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     6001 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     6061 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     6121 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     6181 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     6241 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     6301 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     6361 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     6421 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     6481 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     6541 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     6601 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     6661 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     6721 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     6781 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     6841 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6901 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     6961 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     7021 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     7081 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     7141 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     7201 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7261 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7321 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     7381 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     7441 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     7501 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     7561 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     7621 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     7681 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7741 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt
     7801 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7861 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7921 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7981 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     8041 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     8101 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     8161 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     8221 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     8281 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     8341 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     8401 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     8461 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     8521 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     8581 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     8641 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     8701 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     8761 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8821 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8881 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8941 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     9001 acgtcgacgg atcgggagat ctcccgatcc cctatggt
//
