LOCUS       V045929                 8439 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045929
VERSION     V045929
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        join(8283..8439,1..343)
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             364..963
                     /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"
     misc_feature    977..1565
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             1640..1873
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     promoter        2078..2182
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             2183..3043
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     rep_origin      3214..3802
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    4213..4234
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        4249..4279
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4287..4303
                     /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"
     primer_bind     4311..4327
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     LTR             4588..5221
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    5268..5393
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    5890..6123
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             6308..6352
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    6645..6762
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     enhancer        6814..7117
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        7118..7321
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             7404..8168
                     /label="SNRPA1(NM_003090)"
                     /note="SNRPA1(NM_003090)"
                     /gene="SNRPA1"
     CDS             8199..8264
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
ORIGIN
        1 accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag
       61 gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga
      121 gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt
      181 ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg
      241 cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac
      301 gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca gcccaagctt
      361 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
      421 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
      481 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
      541 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
      601 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
      661 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
      721 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
      781 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
      841 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
      901 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
      961 tgaatcgata gatcctaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     1021 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     1081 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     1141 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     1201 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     1261 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     1321 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     1381 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     1441 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     1501 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     1561 cccgcctgag atcctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     1621 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt
     1681 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     1741 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     1801 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     1861 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     1921 aaatgaatat cagagagtga gaggcccggg ttaattaagg aaagggctag atcattcttg
     1981 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     2041 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2101 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2161 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2221 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2281 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2341 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     2401 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     2461 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     2521 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     2581 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     2641 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     2701 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     2761 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     2821 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2881 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     2941 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3001 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3061 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3121 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3181 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3241 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3301 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3361 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3421 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3481 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3541 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3601 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3661 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3721 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3781 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3841 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3901 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3961 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     4021 gccgattcat taatgcagca agctcatggc tgactaattt tttttattta tgcagaggcc
     4081 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4141 ggcttttgca aaaagctccc cgtggcacga caggtttccc gactggaaag cgggcagtga
     4201 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     4261 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     4321 ctatgacatg attacgaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4381 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa
     4441 ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt
     4501 tcatttactc taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg
     4561 ttaaatatgt actacaaact tagtagttgg aagggctaat tcactcccaa agaagacaag
     4621 atatccttga tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca
     4681 caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag
     4741 ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg
     4801 tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc
     4861 gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat
     4921 atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg
     4981 ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact
     5041 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     5101 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     5161 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     5221 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
     5281 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
     5341 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
     5401 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
     5461 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
     5521 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
     5581 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
     5641 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
     5701 aaaacaaaag taagaccacc gcacagcaag cggccggccg ctgatcttca gacctggagg
     5761 aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga
     5821 accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc
     5881 agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc
     5941 agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca
     6001 gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg
     6061 catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct
     6121 cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc
     6181 tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga
     6241 cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca
     6301 gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg
     6361 gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt
     6421 ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata
     6481 ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg
     6541 aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc
     6601 tcgacggtat cgccgaattc acaaatggca gtattcatcc acaattttaa aagaaaaggg
     6661 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     6721 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
     6781 agcagagatc cagtttggac tagtggagtt ccgcgttaca taacttacgg taaatggccc
     6841 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     6901 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     6961 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     7021 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     7081 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     7141 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     7201 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     7261 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     7321 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
     7381 aagacaccgg cggccgcgcc accatggtca agctgacggc ggagctgatc gagcaggcgg
     7441 cgcagtacac caacgcggtg cgcgaccggg agctggacct ccgggggtat aaaattcccg
     7501 tcattgaaaa tctaggtgct acgttagacc agtttgatgc tattgatttt tctgacaatg
     7561 agatcaggaa actggatggt tttcctttgt tgagaagact gaaaacattg ttagtgaaca
     7621 acaacagaat atgccgtata ggtgagggac ttgatcaggc tctgccctgt ctgacagaac
     7681 tcattctcac caataatagt ctcgtggaac tgggtgatct ggaccctctg gcatctctca
     7741 aatcgctgac ttacctaagt atcctaagaa atccggtaac caataagaag cattacagat
     7801 tgtatgtgat ttataaagtt ccgcaagtca gagtactgga tttccagaaa gtgaaactaa
     7861 aagagcgtca ggaagcagag aaaatgttca agggcaaacg gggtgcacag cttgcaaagg
     7921 atattgccag gagaagcaaa acttttaatc caggtgctgg tttgccaact gacaaaaaga
     7981 aaggtgggcc atctccaggg gatgtagaag caatcaagaa tgccatagca aatgcttcaa
     8041 ctctggctga agtggagagg ctgaaggggt tgctgcagtc tggtcagatc cctggcagag
     8101 aacgcagatc agggcccact gatgatggtg aagaagagat ggaagaagac acagtcacaa
     8161 acgggtccgg tggaggcgga tcaggaggcg gtggatctga ctacaaagac catgacggtg
     8221 attataaaga tcatgacatc gactacaagg atgacgatga caagtagctc gagaattcta
     8281 ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct
     8341 gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag
     8401 gcgccaaccg gctccgttct ttggtggccc cttcgcgcc
//
