LOCUS       V021427                10104 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021427
VERSION     V021427
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             2301..5006
                     /label="Aldh1l1(NM_027406)"
                     /note="Aldh1l1(NM_027406)"
                     /gene="Aldh1l1"
     CDS             5007..5030
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        5074..5285
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5298..5566
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5593..6192
                     /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    6199..6787
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6861..7094
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7166..7287
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7306..7441
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7479..7495)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7503..7519
                     /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(7527..7557)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7572..7593
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7881..8469)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8640..9500)
                     /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(9501..9605)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     10079..10095
                     /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 gagctagcga attcgccacc atgaagattg cagtaatcgg gcagagcctg tttggccagg
     2341 aggtttactg ccagctaagg aaggagggcc atgaggtggt gggtgtgttc accatcccag
     2401 acaaggatgg gaaagcagat cccctgggcc tggaggccga gaaggatggc gtgcccgtgt
     2461 ttaagttccc tcggtggcgg gctcgaggac aggctctgcc cgaggtcgtg gccaagtacc
     2521 aggctctggg tgcagaactc aacgtgctcc ccttctgcag ccagttcata cccatggagg
     2581 tcatcaatgc cccccggcat ggctccatca tctaccatcc atctctgctg cccaggcacc
     2641 gaggggcctc agccatcaac tggaccctca ttcatggaga taagaaagga ggcttcacta
     2701 tcttctgggc tgatgacggt ctggacactg gtgaccttct gctgcagaag gagtgtgacg
     2761 tgcttccaga tgacaccgtg agcacgctgt acaaccgttt cctcttccca gagggcatca
     2821 aagggatggt tcaggctgtg cgactgattg cagagggcac agccccacgc cgtccccagc
     2881 ctgaggaagg agccacctat gagggcattc agaaaaagga gacagccatg atcaactggg
     2941 accagccagc agaggccatt cacaactgga tccgtgggaa tgacaaggtg ccaggtgcct
     3001 ggacagaggc ttgtgggcag aagctgacgt ttttcaactc gacactcaac acttcaggac
     3061 tggtggctca gggagaagct ctacccatcc caggagccca tcgtccaggc ctagtcacca
     3121 aagcgggact catcctcttt gggaatgatg atagaatgtt gctggtgaag aatatccagc
     3181 tggaagatgg caagatgatg ccagcctccc agttcttcaa ggggtctgct agcagtgccc
     3241 tggagctgac cgaggaggag ctggccacag cagaggccgt gcggagctct tggatgcgaa
     3301 ttctgcccaa tgtcccagag gtagaagact ctacagattt cttcaagtca ggagctgcat
     3361 ccgtagatgt tgtgaggctg gtggaggagg tgaaggagct gtgtgacggg ctggagttag
     3421 aaaatgagga tgtttacatg gccaccacct tcggggactt catccagctc ctagtgagga
     3481 agttgagagg ggaggacggc gagagcgagt gtgtcattaa ctacgtggaa aaggcagtga
     3541 agaaactgac tctccaaatg ccctaccagc tcttcatagg cggcgagttt gtggatgctg
     3601 agggcgcgaa gacctacagt accataaacc caacggatgg aagtgtcatc tgccaggtgt
     3661 ctctagctca ggtgagtgat gttgacaagg cggtggcagc agcgaaggag gcctttgaga
     3721 atggactgtg gggaaagata aatgcgcgtg accggggccg gctcctgtac aggttggcgg
     3781 acctcatgga gcagcaccag gaagagctag ccaccattga ggccctggac gcaggtgccg
     3841 tctacacgct ggccctgaag acgcatgtgg gcatgtccat ccagaccttc cgatacttcg
     3901 ctggctggtg tgataagatc cagggtgcca ccatccccat caaccaggct agacccaacc
     3961 gcaacctgac cttgaccaag aaggaacctg ttggggtctg tggtattgtc atcccctgga
     4021 actatccctt aatgatgctg tcctggaaga cagcagcctg cctggctgcc gggaacaccg
     4081 tggtgatcaa gcctgcccag gtgaccccac tcacagcctt gaagtttgca gagctgacac
     4141 tgaaggctgg cattcccaag ggtgtggtca acatcctccc aggatctggc tcgctggttg
     4201 gccagagact ctcagaccac cctgatgtga ggaaaatagg gttcacaggc tccacggagg
     4261 tgggaaaaca catcatgaaa agctgtgccc tgagtaatgt gaagaaggtc tccctggagc
     4321 tgggtggaaa gtcacccctt atcatctttg ctgactgtga cctcaacaaa gctgtgcaga
     4381 tgggcatgag ctccgttttc ttcaacaaag gggagaactg cattgcggca ggccggctct
     4441 ttgtggagga ctccatccat gaccagtttg tgcagaaagt ggtggaggaa gtagggaaga
     4501 tgaaaatcgg caaccccctg gacagagata ccaaccatgg cccgcagaac catgaggccc
     4561 acctgaggaa gctggtggag tattgccaac gtggtgtgaa ggaaggggcc acactggtct
     4621 gtggtgggaa ccaagtccca aggccaggct tcttctttca gccaaccgtt ttcacagacg
     4681 tggaggacca catgtacatc gctaaggagg agtccttcgg gcccatcatg atcatctctc
     4741 ggtttgctga tggggacgtg gatgcagtgc tatctcgggc caatgctaca gaatttggcc
     4801 tggcctctgg tgtcttcact cgggatatca acaaggccct gtatgtcagt gacaaactgc
     4861 aggcgggcac tgtgtttgtc aacacataca acaagaccga tgtggccgca ccttttggag
     4921 gattcaagca gtctggattt ggcaaagacc tgggagaggc ggccctgaat gagtacctgc
     4981 ggatcaagac tgtgactttt gagtacgatt acaaggacga cgatgacaag tagcggccgc
     5041 gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc
     5101 ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg
     5161 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac
     5221 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa
     5281 cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag
     5341 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa
     5401 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc
     5461 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc
     5521 aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg
     5581 cgcctacgct agatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     5641 ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     5701 gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     5761 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     5821 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     5881 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     5941 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     6001 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     6061 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     6121 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     6181 cccggtgcct gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     6241 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     6301 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     6361 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     6421 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     6481 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     6541 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     6601 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     6661 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     6721 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     6781 tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     6841 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat aagatctgct
     6901 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     6961 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     7021 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     7081 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     7141 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
     7201 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     7261 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
     7321 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     7381 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     7441 ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt tcctgtgtga
     7501 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     7561 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     7621 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     7681 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     7741 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     7801 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     7861 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     7921 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     7981 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     8041 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     8101 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     8161 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     8221 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     8281 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     8341 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     8401 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     8461 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     8521 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     8581 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     8641 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     8701 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     8761 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     8821 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     8881 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     8941 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     9001 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     9061 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     9121 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     9181 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     9241 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     9301 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     9361 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     9421 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     9481 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     9541 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     9601 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     9661 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     9721 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     9781 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     9841 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     9901 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
     9961 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
    10021 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
    10081 aaaacgacgg ccagtgccaa gctg
//
