LOCUS       V028332                 7788 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028332
VERSION     V028332
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     repeat_region   1..141
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
                     /rpt_type=inverted
                     /label="AAV2 ITR"
     enhancer        221..524
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        525..727
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     intron          876..1254
                     /note="fusion between human cytomegalovirus intron A and
                     human β-globin intron 2"
                     /label="chimeric intron"
     CDS             1340..2071
                     /label="Klf9(NM_010638)"
                     /note="Klf9(NM_010638)"
                     /gene="Klf9"
     CDS             2078..2785
                     /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
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEGYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             2792..2845
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             2849..4498
                     /EC_number=""
                     /codon_start=1
                     /gene="luc+"
                     /note="enhanced&nbsp;luc+ version of the luciferase gene"
                     /product="firefly luciferase"
                     /protein_id=""
                     /transl_table=1
                     /translation="EDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAH
                     IEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPA
                     NDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSM
                     YTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHAR
                     DPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKI
                     QSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYG
                     LTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSG
                     YVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILL
                     QHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVF
                     VDEVPKGLTGKLDARKIREILIKAKKGGKIAV*"
                     /label="luciferase"
     polyA_signal    4535..5011
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     repeat_region   5051..5191
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
                     /rpt_type=inverted
                     /label="AAV2 ITR"
     rep_origin      5266..5721
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6003..6107
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             6108..6968
                     /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"
     rep_origin      7139..7727
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtggagc tagttattaa tagtaatcaa
      181 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      241 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      301 ttcccatagt aacgtcaata gggactttcc attgacgtca atgggtggag tatttacggt
      361 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      421 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      481 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      541 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      601 ttgacgtcaa tgggagtttg ttttgcacca aaatcaacgg gactttccaa aatgtcgtaa
      661 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      721 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
      781 ccatagaaga caccgggacc gatccagcct ccgcggattc gaatcccggc cgggaacggt
      841 gcattggaac gcggattccc cgtgccaaga gtgacgtaag taccgcctat agagtctata
      901 ggcccacaaa aaatgctttc ttcttttaat atactttttt gtttatctta tttctaatac
      961 tttccctaat ctctttcttt cagggcaata atgatacaat gtatcatgcc tctttgcacc
     1021 attctaaaga ataacagtga taatttctgg gttaaggcaa tagcaatatt tctgcatata
     1081 aatatttctg catataaatt gtaactgatg taagaggttt catattgcta atagcagcta
     1141 caatccagct accattctgc ttttatttta tggttgggat aaggctggat tattctgagt
     1201 ccaagctagg cccttttgct aatcatgttc atacctctta tcttcctccc acagctcctg
     1261 ggcaacgtgc tggtctgtgt gctggcccat cactttggca aagaattggg attcgaacat
     1321 cgattgaatt ccagccacca tgtccgcggc cgcctacatg gacttcgtgg ctgcccagtg
     1381 tctggtttcc atctccaacc gcgccgccgt gccggagcac gggggcgctc cggaagccga
     1441 gcggctgcga ctacctgagc gcgaggtgac caaggaacac ggtgacccgg gggacacctg
     1501 gaaggattat tgcacgctgg tcactatcgc caagagcttg ttggacctca acaaataccg
     1561 acccatccag accccctcgg tgtgcagcga cagtctggag agtcccgatg aggatatagg
     1621 atccgacagc gacgtgacca ccgaatctgg gtcgagtcct tcccacagcc cggaggagag
     1681 acaggattct ggcagcgcgc ccagcccact ctccctcctc cactctggag tggcttcgaa
     1741 ggggaaacac gcctccgaaa agaggcacaa gtgcccctac agtggctgtg ggaaagtcta
     1801 tggaaaatcc tcccatctta aagcccatta cagagtgcat acaggtgaac ggccctttcc
     1861 ctgcacgtgg ccagactgcc ttaaaaagtt ctcgcgctcg gatgagctga cccgccacta
     1921 ccggacccac actggggaaa agcagttccg ttgcccactg tgtgagaaga gattcatgag
     1981 gagtgaccat ctcaccaagc atgcccggcg tcacaccgag ttccatccca gcatgatcaa
     2041 gagatcaaaa aaggctcttg ccagcccctt gggatccatg gtgagcaagg gcgaggagga
     2101 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
     2161 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     2221 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
     2281 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
     2341 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     2401 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     2461 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     2521 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
     2581 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
     2641 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
     2701 ccacaacgag ggctacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
     2761 cggcggcatg gacgagctgt acaagccccg ggagggcaga ggaagtcttc taacatgcgg
     2821 tgacgtggag gagaatcccg gccctcgaga agacgccaaa aacataaaga aaggcccggc
     2881 gccattctat ccgctggaag atggaaccgc tggagagcaa ctgcataagg ctatgaagag
     2941 atacgccctg gttcctggaa caattgcttt tacagatgca catatcgagg tggacatcac
     3001 ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac gatatgggct
     3061 gaatacaaat cacagaatcg tcgtatgcag tgaaaactct cttcaattct ttatgccggt
     3121 gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg aacgacattt ataatgaacg
     3181 tgaattgctc aacagtatgg gcatttcgca gcctaccgtg gtgttcgttt ccaaaaaggg
     3241 gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc atccaaaaaa ttattatcat
     3301 ggattctaaa acggattacc agggatttca gtcgatgtac acgttcgtca catctcatct
     3361 acctcccggt tttaatgaat acgattttgt gccagagtcc ttcgataggg acaagacaat
     3421 tgcactgatc atgaactcct ctggatctac tggtctgcct aaaggtgtcg ctctgcctca
     3481 tagaactgcc tgcgtgagat tctcgcatgc cagagatcct atttttggca atcaaatcat
     3541 tccggatact gcgattttaa gtgttgttcc attccatcac ggttttggaa tgtttactac
     3601 actcggatat ttgatatgtg gatttcgagt cgtcttaatg tatagatttg aagaagagct
     3661 gtttctgagg agccttcagg attacaagat tcaaagtgcg ctgctggtgc caaccctatt
     3721 ctccttcttc gccaaaagca ctctgattga caaatacgat ttatctaatt tacacgaaat
     3781 tgcttctggt ggcgctcccc tctctaagga agtcggggaa gcggttgcca agaggttcca
     3841 tctgccaggt atcaggcaag gatatgggct cactgagact acatcagcta ttctgattac
     3901 gcccgagggg gatgataaac cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa
     3961 ggttgtggat ctggataccg ggaaaacgct gggcgttaat caaagaggcg aactgtgtgt
     4021 gagaggtcct atgattatgt ccggttatgt aaacaatccg gaagcgacca acgccttgat
     4081 tgacaaggat ggatggctac attctggaga catagcttac tgggacgaag acgaacactt
     4141 cttcatcgtt gaccgcctga agtctctgat taagtacaaa ggctatcagg tggctcccgc
     4201 tgaattggaa tccatcttgc tccaacaccc caacatcttc gacgcaggtg tcgcaggtct
     4261 tcccgacgat gacgccggtg aacttcccgc cgccgttgtt gttttggagc acggaaagac
     4321 gatgacggaa aaagagatcg tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt
     4381 gcgcggagga gttgtgtttg tggacgaagt accgaaaggt cttaccggaa aactcgacgc
     4441 aagaaaaatc agagagatcc tcataaaggc caagaagggc ggaaagatcg ccgtgtaaaa
     4501 gcttgcctcg agcagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc
     4561 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa
     4621 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg
     4681 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg
     4741 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt
     4801 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag
     4861 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt
     4921 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca
     4981 ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc
     5041 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     5101 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     5161 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     5221 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     5281 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     5341 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     5401 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     5461 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     5521 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     5581 ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg attttgccga
     5641 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     5701 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     5761 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     5821 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     5881 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     5941 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     6001 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6061 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6121 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6181 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     6241 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     6301 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     6361 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     6421 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     6481 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     6541 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     6601 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     6661 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     6721 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     6781 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     6841 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6901 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6961 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7021 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7081 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7141 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     7201 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     7261 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     7321 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     7381 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     7441 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     7501 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     7561 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     7621 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     7681 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     7741 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
