LOCUS       V036196                 9190 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036196
VERSION     V036196
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             2302..4044
                     /codon_start=1
                     /product="human PTEN-induced putative kinase 1, a
                     mitochondria serine/threonine protein kinase"
                     /transl_table=1
                     /translation="MAVRQALGRGLQLGRALLLRFTGKPGRAYGLGRPGPAAGCVRGER
                     PGWAAGPGAEPRRVGLGLPNRLRFFRQSVAGLAARLQRQFVVRAWGCAGPCGRAVFLAF
                     GLGLGLIEEKQAESRRAVSACQEIQAIFTQKSKPGPDPLDTRRLQGFRLEEYLIGQSIG
                     KGCSAAVYEATMPTLPQNLEVTKSTGLLPGRGPGTSAPGEGQERAPGAPAFPLAIKMMW
                     NISAGSSSEAILNTMSQELVPASRVALAGEYGAVTYRKSKRGPKQLAPHPNIIRVLRAF
                     TSSVPLLPGALVDYPDVLPSRLHPEGLGHGRTLFLVMKNYPCTLRQYLCVNTPSPRLAA
                     MMLLQLLEGVDHLVQQGIAHRDLKSDNILVELDPDGCPWLVIADFGCCLADESIGLQLP
                     FSSWYVDRGGNGCLMAPEVSTARPGPRAVIDYSKADAWAVGAIAYEIFGLVNPFYGQGK
                     AHLESRSYQEAQLPALPESVPPDVRQLVRALLQREASKRPSARVAANVLHLSLWGEHIL
                     ALKNLKLDKMVGWLLQQSAATLLANRLTEKCCVETKMKMLFLANLECETLCQAALLLCS
                     WRAAL"
                     /label="PINK1"
     CDS             4051..4116
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4160..4371
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4384..4652
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4679..5278
                     /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    5285..5873
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5947..6180
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6252..6373
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6392..6527
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6565..6581)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6589..6605
                     /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(6613..6643)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6658..6679
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6967..7555)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7726..8586)
                     /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(8587..8691)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9165..9181
                     /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 agagctagcg aattcgccac catggcggtg cgacaggcgc tgggccgcgg cctgcagctg
     2341 ggtcgagcgc tgctgctgcg cttcacgggc aagcccggcc gggcctacgg cttggggcgg
     2401 ccgggcccgg cggcgggctg tgtccgcggg gagcgtccag gctgggccgc aggaccgggc
     2461 gcggagcctc gcagggtcgg gctcgggctc cctaaccgtc tccgcttctt ccgccagtcg
     2521 gtggccgggc tggcggcgcg gttgcagcgg cagttcgtgg tgcgggcctg gggctgcgcg
     2581 ggcccttgcg gccgggcagt ctttctggcc ttcgggctag ggctgggcct catcgaggaa
     2641 aaacaggcgg agagccggcg ggcggtctcg gcctgtcagg agatccaggc aatttttacc
     2701 cagaaaagca agccggggcc tgacccgttg gacacgagac gcttgcaggg ctttcggctg
     2761 gaggagtatc tgatagggca gtccattggt aagggctgca gtgctgctgt gtatgaagcc
     2821 accatgccta cattgcccca gaacctggag gtgacaaaga gcaccgggtt gcttccaggg
     2881 agaggcccag gtaccagtgc accaggagaa gggcaggagc gagctccggg ggcccctgcc
     2941 ttccccttgg ccatcaagat gatgtggaac atctcggcag gttcctccag cgaagccatc
     3001 ttgaacacaa tgagccagga gctggtccca gcgagccgag tggccttggc tggggagtat
     3061 ggagcagtca cttacagaaa atccaagaga ggtcccaagc aactagcccc tcaccccaac
     3121 atcatccggg ttctccgcgc cttcacctct tccgtgccgc tgctgccagg ggccctggtc
     3181 gactaccctg atgtgctgcc ctcacgcctc caccctgaag gcctgggcca tggccggacg
     3241 ctgttcctcg ttatgaagaa ctatccctgt accctgcgcc agtacctttg tgtgaacaca
     3301 cccagccccc gcctcgccgc catgatgctg ctgcagctgc tggaaggcgt ggaccatctg
     3361 gttcaacagg gcatcgcgca cagagacctg aaatccgaca acatccttgt ggagctggac
     3421 ccagacggct gcccctggct ggtgatcgca gattttggct gctgcctggc tgatgagagc
     3481 atcggcctgc agttgccctt cagcagctgg tacgtggatc ggggcggaaa cggctgtctg
     3541 atggccccag aggtgtccac ggcccgtcct ggccccaggg cagtgattga ctacagcaag
     3601 gctgatgcct gggcagtggg agccatcgcc tatgaaatct tcgggcttgt caatcccttc
     3661 tacggccagg gcaaggccca ccttgaaagc cgcagctacc aagaggctca gctacctgca
     3721 ctgcccgagt cagtgcctcc agacgtgaga cagttggtga gggcactgct ccagcgagag
     3781 gccagcaaga gaccatctgc ccgagtagcc gcaaatgtgc ttcatctaag cctctggggt
     3841 gaacatattc tagccctgaa gaatctgaag ttagacaaga tggttggctg gctcctccaa
     3901 caatcggccg ccactttgtt ggccaacagg ctcacagaga agtgttgtgt ggaaacaaaa
     3961 atgaagatgc tctttctggc taacctggag tgtgaaacgc tctgccaggc agccctcctc
     4021 ctctgctcat ggagggcagc cctgggatcc gactacaaag accatgacgg tgattataaa
     4081 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat
     4141 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
     4201 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
     4261 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
     4321 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt
     4381 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc
     4441 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc
     4501 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta
     4561 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt
     4621 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagat
     4681 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg
     4741 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg
     4801 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     4861 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     4921 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     4981 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     5041 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     5101 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     5161 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     5221 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagt
     5281 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     5341 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     5401 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     5461 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     5521 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     5581 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     5641 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     5701 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     5761 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     5821 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac
     5881 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     5941 ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg
     6001 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     6061 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     6121 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     6181 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     6241 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6301 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6361 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc
     6421 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     6481 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag
     6541 agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6601 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6661 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6721 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6781 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6841 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6901 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6961 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7021 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7081 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7141 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7201 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7261 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7321 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7381 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     7441 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7501 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7561 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7621 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7681 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7741 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7801 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7861 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7921 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7981 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     8041 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8101 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8161 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8221 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8281 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8341 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8401 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8461 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8521 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8581 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8641 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8701 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     8761 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     8821 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     8881 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     8941 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     9001 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     9061 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     9121 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     9181 tgccaagctg
//
