LOCUS       V021774                 9635 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021774
VERSION     V021774
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /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    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2293..3675
                     /label="RNH1(NM_002939)"
                     /note="RNH1(NM_002939)"
                     /gene="RNH1"
     CDS             3733..4479
                     /codon_start=1
                     /note="more active than the original APEX due to the A134P
                     mutation"
                     /product="soybean ascorbate peroxidase, engineered to serve
                     as a monomeric tag for electron microscopy and live-cell
                     proteomics (Lam et al., 2015)"
                     /transl_table=1
                     /translation="GKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAGT
                     FDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVAV
                     EVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGHT
                     IGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVDK
                     YAADEDAFFADYAEAHQKLSELGFADA"
                     /label="APEX2"
     CDS             4489..4554
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4605..4816
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4829..5097
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5124..5723
                     /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    5730..6318
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6392..6625
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6697..6818
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6837..6972
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7010..7026)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7034..7050
                     /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(7058..7088)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7103..7124
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7412..8000)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8171..9031)
                     /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(9032..9136)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9610..9626
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga 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 ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta
     2281 gagctagcca ccatgagcct ggacatccag agcctggaca tccagtgtga ggagctgagc
     2341 gacgctagat gggccgagct cctccctctg ctccagcagt gccaagtggt caggctggac
     2401 gactgtggcc tcacggaagc acggtgcaag gacatcagct ctgcacttcg agtcaaccct
     2461 gcactggcag agctcaacct gcgcagcaac gagctgggcg atgtcggcgt gcattgcgtg
     2521 ctccagggcc tgcagacccc ctcctgcaag atccagaagc tgagcctcca gaactgctgc
     2581 ctgacggggg ccggctgcgg ggtcctgtcc agcacactac gcaccctgcc caccctgcag
     2641 gagctgcacc tcagcgacaa cctcttgggg gatgcgggcc tgcagctgct ctgcgaagga
     2701 ctcctggacc cccagtgccg cctggaaaag ctgcagctgg agtattgcag cctctcggct
     2761 gccagctgcg agcccctggc ctccgtgctc agggccaagc cggacttcaa ggagctcacg
     2821 gttagcaaca acgacatcaa tgaggctggc gtccgtgtgc tgtgccaggg cctgaaggac
     2881 tccccctgcc agctggaggc gctcaagctg gagagctgcg gtgtgacatc agacaactgc
     2941 cgggacctgt gcggcattgt ggcctccaag gcctcgctgc gggagctggc cctgggcagc
     3001 aacaagctgg gtgatgtggg catggcggag ctgtgcccag ggctgctcca ccccagctcc
     3061 aggctcagga ccctgtggat ctgggagtgt ggcatcactg ccaagggctg cggggatctg
     3121 tgccgtgtcc tcagggccaa ggagagcctg aaggagctca gcctggccgg caacgagctg
     3181 ggggatgagg gtgcccgact gctgtgtgag accctgctgg aacctggctg ccagctggag
     3241 tcgctgtggg tgaagtcctg cagcttcaca gccgcctgct gctcccactt cagctcagtg
     3301 ctggcccaga acaggtttct cctggagcta cagataagca acaacaggct ggaggatgcg
     3361 ggcgtgcggg agctgtgcca gggcctgggc cagcctggct ctgtgctgcg ggtgctctgg
     3421 ttggccgact gcgatgtgag tgacagcagc tgcagcagcc tcgccgcaac cctgttggcc
     3481 aaccacagcc tgcgtgagct ggacctcagc aacaactgcc tgggggacgc cggcatcctg
     3541 cagctggtgg agagcgtccg gcagccgggc tgcctcctgg agcagctggt cctgtacgac
     3601 atttactggt ctgaggagat ggaggaccgg ctgcaggccc tggagaagga caagccatcc
     3661 ctgagggtca tctccctcga ggggggtgga ggtggcggga gcggtggggg tggcagcgga
     3721 ggtggtggga gcggaaagtc ttacccaact gtgagtgctg attaccagga cgccgttgag
     3781 aaggcgaaga agaagctcag aggcttcatc gctgagaaga gatgcgctcc tctaatgctc
     3841 cgtttggcat tccactctgc tggaaccttt gacaagggca cgaagaccgg tggacccttc
     3901 ggaaccatca agcaccctgc cgaactggct cacagcgcta acaacggtct tgacatcgct
     3961 gttaggcttt tggagccact caaggcggag ttccctattt tgagctacgc cgatttctac
     4021 cagttggctg gcgttgttgc cgttgaggtc acgggtggac ctaaggttcc attccaccct
     4081 ggaagagagg acaagcctga gccaccacca gagggtcgct tgcccgatcc cactaagggt
     4141 tctgaccatt tgagagatgt gtttggcaaa gctatggggc ttactgacca agatatcgtt
     4201 gctctatctg ggggtcacac tattggagct gcacacaagg agcgttctgg atttgagggt
     4261 ccctggacct ctaatcctct tattttcgac aactcatact tcacggagtt gttgagtggt
     4321 gagaaggaag gtctccttca gctaccttct gacaaggctc ttttgtctga ccctgtattc
     4381 cgccctctcg ttgataaata tgcagcggac gaagatgcct tctttgctga ttacgctgag
     4441 gctcaccaaa agctttccga gcttgggttt gctgatgccc gcggccgcga ctacaaagac
     4501 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtgagga
     4561 tccgcggccg cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc
     4621 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag
     4681 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     4741 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     4801 tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg
     4861 ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg
     4921 tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct
     4981 ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac
     5041 cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa
     5101 gctgtgaccg gcgcctacgc tagatgaccg agtacaagcc cacggtgcgc ctcgccaccc
     5161 gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
     5221 cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct
     5281 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
     5341 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
     5401 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
     5461 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg
     5521 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
     5581 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
     5641 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
     5701 tgacccgcaa gcccggtgcc tgagtcgaca atcaacctct ggattacaaa atttgtgaaa
     5761 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     5821 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     5881 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     5941 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     6001 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     6061 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     6121 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     6181 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     6241 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     6301 tttgggccgc ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat
     6361 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaaa
     6421 taagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     6481 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     6541 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     6601 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     6661 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     6721 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6781 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     6841 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6901 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     6961 tttttggagg cctagacttt tgcagagacg gcccaaattc gtaatcatgg tcatagctgt
     7021 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     7081 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     7141 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     7201 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     7261 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     7321 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     7381 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     7441 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     7501 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     7561 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     7621 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7681 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7741 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7801 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     7861 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7921 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7981 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     8041 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     8101 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     8161 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     8221 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     8281 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     8341 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     8401 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     8461 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     8521 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     8581 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     8641 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8701 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8761 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8821 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8881 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8941 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     9001 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     9061 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     9121 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     9181 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     9241 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     9301 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     9361 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     9421 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
     9481 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     9541 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     9601 cacgacgttg taaaacgacg gccagtgcca agctg
//
