LOCUS       V044150                 7889 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044150
VERSION     V044150
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             908..2848
                     /label="Tnip1(NM_021327)"
                     /note="Tnip1(NM_021327)"
                     /gene="Tnip1"
     CDS             2897..3607
                     /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"
     polyA_signal    3651..3875
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3921..4349
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4363..4692
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4759..5358
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    5532..5653
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5702..5718)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5726..5742
                     /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(5750..5780)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5795..5816
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6104..6689)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6860..7720)
                     /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(7721..7825)
                     /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 ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gaagggagag gaccctacgg gatctacgac ccagggggca gcacgcctct
      961 gggagaggtg tccgcagctt ttgaacgtct agtggaggag aatactcggc tgaagggaaa
     1021 aatgcaaggg ataaagatgt taggggagct tctggaggag tctcagatgg aagcgtccag
     1081 actccggcag aaggcagagg agctggtcaa ggacagcgag ctgtcaccac cgacatctgc
     1141 cccctccttg gtctcctttg atgacctggc tgagctcaca ggacaggata caaaggtcca
     1201 ggtacatcct gctaccagca ctgccgccac caccaccgcc accgccacca cgggaaactc
     1261 catggagaag cccgagccag cctccaaatc tccgtccaat ggcgcctcct cggactttga
     1321 agtggtccct actgaggagc agaattcacc cgaaactggc agccacccta cgaacatgat
     1381 ggacctgggg cccccacccc cagaggacag caacctgaag ctccacctgc agcgcctgga
     1441 gaccaccctt agcgtgtgtg cagaggagcc agaccacagc cagctcttca cccacctggg
     1501 ccgcatggcc ctcgagttca acaggttggc ctccaaagtg cataaaaatg agcagcgcac
     1561 ctccatcctg cagaccttat gtgagcagct gcgccaggag aatgaagccc tgaaggccaa
     1621 gctggacaag ggcctggaac agcgggatct ggctgctgag aggctgcggg aggaaaacac
     1681 ggagctcaag aaactgttga tgaacagcag ctgcaaagag ggactctgtg ggcagcccag
     1741 ctccccaaag ccagagggtg ctggcaagaa gggcgtggct ggacagcagc aggccagtgt
     1801 gatggcgagt aaagtccctg aagcgggggc ctttggagca gctgagaaga aggtgaagtt
     1861 gctagaacag caacgcatgg agctgctgga agtgaacaag cagtgggacc agcatttccg
     1921 gtccatgaag cagcagtatg agcagaagat cacagagctt cgccagaagc tggtggacct
     1981 gcagaaacag gtaactgagc tggaggccga acgggagcag aagcagcgtg actttgaccg
     2041 gaaactcctc ctggccaaat cgaagataga gatggaagag accgacaagg agcagctgac
     2101 agcagaggcc aaggaactgc gccagaaggt caggtaccta caggatcagc tgagcccgct
     2161 cacaaggcaa cgagaatacc aggagaagga gatccagcgg ctcaataagg ccctggagga
     2221 ggccctcagc atccaggcct ctccatcatc tccgcctgca gcttttggga gtccagaagg
     2281 cgttgggggc catctgagga agcaggaact agtgacacag aatgagttgc tgaaacagca
     2341 ggtaaagatc tttgaagagg acttccagag ggaacggagt gaccgtgaac gcatgaatga
     2401 agagaaggag gagctgaaga agcaagtaga gaagctgcag gcccaggtca ccctgactaa
     2461 tgcccagctc aaaactctca aagaggagga gaaggccaag gaagccctca aacagcagaa
     2521 gaggaaagca aaggcttcgg gagagcgcta ccacatggaa ccccaccctg agcacgtctg
     2581 cggcgcctat ccctatgcct acccacccat gccagccatg gtacctcacc atgcctacaa
     2641 ggactggtcc cagatccgat accctccacc ccctgtgccc atggagcacc cgcccccaca
     2701 ccccaactct cgcctcttcc atctgccgga gtacacctgg cgtccaccct gtgcagggat
     2761 tcggaatcag agctctcaag tgatggaccc gcccccagac aggcctgcag agccagagtc
     2821 tgcagacaat gactgtgatg ggccccagga attcgtcatg gatatcggtg gaggcggatc
     2881 aggaggcggt ggatctatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
     2941 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
     3001 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     3061 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
     3121 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
     3181 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     3241 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     3301 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat
     3361 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     3421 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     3481 cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
     3541 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
     3601 caagtagctc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     3661 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3721 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3781 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3841 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     3901 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3961 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4021 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4081 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4141 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4201 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4261 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4321 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     4381 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4441 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4501 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4561 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4621 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4681 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     4741 ggatgaggat cgtttcgcat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     4801 gacgtccccc gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     4861 cacaccgtcg acccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     4921 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     4981 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     5041 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     5101 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     5161 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     5221 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     5281 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     5341 cgcaagcccg gtgcctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5401 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5461 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5521 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5581 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5641 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     5701 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5761 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5821 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5881 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5941 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6001 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6061 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6121 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6181 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6241 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6301 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6361 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6421 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6481 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6541 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6601 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc
     6661 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6721 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6781 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6841 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6901 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6961 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7021 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7081 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7141 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7201 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7261 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7321 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7381 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7441 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7501 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7561 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7621 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7681 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7741 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7801 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     7861 gatcgggaga tctcccgatc ccctatggt
//
