LOCUS       V028374                 8892 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028374
VERSION     V028374
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        3..229
                     /note="Rous sarcoma virus enhancer/promoter"
                     /label="RSV promoter"
     LTR             230..410
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    457..582
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1075..1308
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /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    1803..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     protein_bind    1938..1958
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of attB"
                     /note="core recombination site for the Gateway® BP
                     reaction"
                     /label="attB4"
     enhancer        2019..2322
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2323..2526
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2566..3561
                     /label="ADORA2B(NM_000676)"
                     /note="ADORA2B(NM_000676)"
                     /gene="ADORA2B"
     misc_feature    3607..4195
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     promoter        4226..4725
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4742..5341
                     /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"
     LTR             5413..5646
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5718..5839
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5879..6014
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(6035..6053)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(6063..6079)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      6221..6676
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6702..6806
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             6807..7667
                     /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      7838..8426
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    8714..8735
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        8750..8780
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8788..8804
                     /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     8812..8828
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        8849..8867
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /label="T3 promoter"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgcta
     1801 gcttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
     1861 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt
     1921 actagtgatt atcggatcaa ctttgtatag aaaagttgta gttattaata gtaatcaatt
     1981 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     2041 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     2101 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     2161 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     2221 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     2281 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     2341 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     2401 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     2461 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     2521 agagctggtt tagtgaaccg tcagatccaa gttggatccg ccaccatgct gctggagaca
     2581 caggacgcgc tgtacgtggc gctggagctg gtcatcgccg cgctttcggt ggcgggcaac
     2641 gtgctggtgt gcgccgcggt gggcacggcg aacactctgc agacgcccac caactacttc
     2701 ctggtgtccc tggctgcggc cgacgtggcc gtggggctct tcgccatccc ctttgccatc
     2761 accatcagcc tgggcttctg cactgacttc tacggctgcc tcttcctcgc ctgcttcgtg
     2821 ctggtgctca cgcagagctc catcttcagc cttctggccg tggcagtcga cagatacctg
     2881 gccatctgtg tcccgctcag gtataaaagt ttggtcacgg ggacccgagc aagaggggtc
     2941 attgctgtcc tctgggtcct tgcctttggc atcggattga ctccattcct ggggtggaac
     3001 agtaaagaca gtgccaccaa caactgcaca gaaccctggg atggaaccac gaatgaaagc
     3061 tgctgccttg tgaagtgtct ctttgagaat gtggtcccca tgagctacat ggtatatttc
     3121 aatttctttg ggtgtgttct gcccccactg cttataatgc tggtgatcta cattaagatc
     3181 ttcctggtgg cctgcaggca gcttcagcgc actgagctga tggaccactc gaggaccacc
     3241 ctccagcggg agatccatgc agccaagtca ctggccatga ttgtggggat ttttgccctg
     3301 tgctggttac ctgtgcatgc tgttaactgt gtcactcttt tccagccagc tcagggtaaa
     3361 aataagccca agtgggcaat gaatatggcc attcttctgt cacatgccaa ttcagttgtc
     3421 aatcccattg tctatgctta ccggaaccga gacttccgct acacttttca caaaattatc
     3481 tccaggtatc ttctctgcca agcagatgtc aagagtggga atggtcaggc tggggtacag
     3541 cctgctctcg gtgtgggcct atgactcgag tctagatgta caaagtggtg ataatcgaat
     3601 tccgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     3661 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     3721 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     3781 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     3841 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     3901 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     3961 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     4021 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     4081 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     4141 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcgg
     4201 gaattcccgc ggttcgaatt ctaccgggta ggggaggcgc ttttcccaag gcagtctgga
     4261 gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc
     4321 acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg
     4381 ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc
     4441 aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct
     4501 gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct
     4561 ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg cgggctcagg
     4621 ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc
     4681 acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctc acgtggccac
     4741 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     4801 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     4861 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     4921 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     4981 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     5041 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     5101 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     5161 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     5221 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     5281 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     5341 aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     5401 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt
     5461 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     5521 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     5581 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     5641 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
     5701 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5761 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5821 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat
     5881 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5941 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     6001 cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc
     6061 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     6121 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     6181 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
     6241 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     6301 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     6361 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     6421 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     6481 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     6541 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     6601 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     6661 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     6721 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     6781 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     6841 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     6901 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     6961 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     7021 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     7081 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     7141 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     7201 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     7261 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     7321 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     7381 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     7441 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     7501 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     7561 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     7621 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     7681 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     7741 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     7801 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     7861 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     7921 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     7981 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     8041 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     8101 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     8161 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     8221 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     8281 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     8341 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     8401 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     8461 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     8521 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     8581 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     8641 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     8701 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     8761 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     8821 gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg
     8881 agctgcaagc tt
//
