LOCUS       V033835                 9776 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033835
VERSION     V033835
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             2290..3039
                     /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="MGKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAG
                     TFDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVA
                     VEVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGH
                     TIGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVD
                     KYAADEDAFFADYAEAHQKLSELGFADA"
                     /label="APEX2"
     CDS             3040..4269
                     /label="LAMP2(NM_002294)"
                     /note="LAMP2(NM_002294)"
                     /gene="LAMP2"
     promoter        4319..4530
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4543..4811
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4839..5864
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /note="confers resistance to hygromycin"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /transl_table=1
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E*"
                     /label="HygR"
     misc_feature    5871..6459
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6533..6766
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6838..6959
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6978..7113
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7151..7167)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7175..7191
                     /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(7199..7229)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7244..7265
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7553..8141)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8312..9172)
                     /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(9173..9277)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9751..9767
                     /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 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagccacca tgggaaagtc ttacccaact gtgagtgctg attaccagga cgccgttgag
     2341 aaggcgaaga agaagctcag aggcttcatc gctgagaaga gatgcgctcc tctaatgctc
     2401 cgtttggcat tccactctgc tggaaccttt gacaagggca cgaagaccgg tggacccttc
     2461 ggaaccatca agcaccctgc cgaactggct cacagcgcta acaacggtct tgacatcgct
     2521 gttaggcttt tggagccact caaggcggag ttccctattt tgagctacgc cgatttctac
     2581 cagttggctg gcgttgttgc cgttgaggtc acgggtggac ctaaggttcc attccaccct
     2641 ggaagagagg acaagcctga gccaccacca gagggtcgct tgcccgatcc cactaagggt
     2701 tctgaccatt tgagagatgt gtttggcaaa gctatggggc ttactgacca agatatcgtt
     2761 gctctatctg ggggtcacac tattggagct gcacacaagg agcgttctgg atttgagggt
     2821 ccctggacct ctaatcctct tattttcgac aactcatact tcacggagtt gttgagtggt
     2881 gagaaggaag gtctccttca gctaccttct gacaaggctc ttttgtctga ccctgtattc
     2941 cgccctctcg ttgacaaata tgcagcggac gaagatgcct tctttgctga ttacgctgag
     3001 gctcaccaaa agctttccga gcttgggttt gctgatgcca tggtgtgctt ccgcctcttc
     3061 ccggttccgg gctcagggct cgttctggtc tgcctagtcc tgggagctgt gcggtcttat
     3121 gcattggaac ttaatttgac agattcagaa aatgccactt gcctttatgc aaaatggcag
     3181 atgaatttca cagtacgcta tgaaactaca aataaaactt ataaaactgt aaccatttca
     3241 gaccatggca ctgtgacata taatggaagc atttgtgggg atgatcagaa tggtcccaaa
     3301 atagcagtgc agttcggacc tggcttttcc tggattgcga attttaccaa ggcagcatct
     3361 acttattcaa ttgacagcgt ctcattttcc tacaacactg gtgataacac aacatttcct
     3421 gatgctgaag ataaaggaat tcttactgtt gatgaacttt tggccatcag aattccattg
     3481 aatgaccttt ttagatgcaa tagtttatca actttggaaa agaatgatgt tgtccaacac
     3541 tactgggatg ttcttgtaca agcttttgtc caaaatggca cagtgagcac aaatgagttc
     3601 ctgtgtgata aagacaaaac ttcaacagtg gcacccacca tacacaccac tgtgccatct
     3661 cctactacaa cacctactcc aaaggaaaaa ccagaagctg gaacctattc agttaataat
     3721 ggcaatgata cttgtctgct ggctaccatg gggctgcagc tgaacatcac tcaggataag
     3781 gttgcttcag ttattaacat caaccccaat acaactcact ccacaggcag ctgccgttct
     3841 cacactgctc tacttagact caatagcagc accattaagt atctagactt tgtctttgct
     3901 gtgaaaaatg aaaaccgatt ttatctgaag gaagtgaaca tcagcatgta tttggttaat
     3961 ggctccgttt tcagcattgc aaataacaat ctcagctact gggatgcccc cctgggaagt
     4021 tcttatatgt gcaacaaaga gcagactgtt tcagtgtctg gagcatttca gataaatacc
     4081 tttgatctaa gggttcagcc tttcaatgtg acacaaggaa agtattctac agctcaagac
     4141 tgcagtgcag atgacgacaa cttccttgtg cccatagcgg tgggagctgc cttggcagga
     4201 gtacttattc tagtgttgct ggcttatttt attggtctca agcaccatca tgctggatat
     4261 gagcaatttt agggatccgc ggccgcaagg atctgcgatc gctccggtgc ccgtcagtgg
     4321 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacg
     4381 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     4441 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     4501 tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct
     4561 tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg
     4621 cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg
     4681 tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc
     4741 acgctttgcc tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc
     4801 cgttacagat ccaagctgtg accggcgcct acgtcgacat gaaaaagcct gaactcaccg
     4861 cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc
     4921 tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc
     4981 tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg
     5041 catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc gagagcctga
     5101 cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac
     5161 tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta
     5221 gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc
     5281 gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg
     5341 acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact
     5401 gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca
     5461 atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg
     5521 aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct
     5581 acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc
     5641 gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt
     5701 gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac
     5761 aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata
     5821 gtggaaaccg acgccccagc actcgtccga gggcaaagga ataggtcgac aatcaacctc
     5881 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     5941 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     6001 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     6061 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     6121 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     6181 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     6241 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     6301 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     6361 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     6421 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggtaccttt aagaccaatg
     6481 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     6541 ctaattcact cccaacgaaa ataagatctg ctttttgctt gtactgggtc tctctggtta
     6601 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     6661 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     6721 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt
     6781 catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac
     6841 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     6901 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     6961 catgtctggc tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     7021 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     7081 tccagaagta gtgaggaggc ttttttggag gcctagactt ttgcagagac ggcccaaatt
     7141 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     7201 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     7261 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     7321 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     7381 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     7441 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     7501 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7561 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7621 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     7681 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     7741 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     7801 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     7861 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     7921 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     7981 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     8041 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     8101 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     8161 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     8221 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     8281 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     8341 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     8401 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     8461 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     8521 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     8581 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     8641 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     8701 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     8761 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     8821 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     8881 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     8941 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     9001 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     9061 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     9121 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     9181 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     9241 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     9301 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     9361 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     9421 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     9481 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     9541 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     9601 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     9661 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     9721 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg
//
