LOCUS       V042700                 9064 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042700
VERSION     V042700
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..3918
                     /label="ALDH7A1(NM_001182)"
                     /note="ALDH7A1(NM_001182)"
                     /gene="ALDH7A1"
     CDS             3925..3990
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4034..4245
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4258..4526
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4553..5152
                     /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    5159..5747
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5821..6054
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6126..6247
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6266..6401
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6439..6455)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6463..6479
                     /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(6487..6517)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6532..6553
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6841..7429)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7600..8460)
                     /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(8461..8565)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9039..9055
                     /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 catgtggcgc cttcctcgcg cgctgtgtgt gcacgctgca
     2341 aagaccagca agctctctgg accttggagc aggcctgccg ccttcatgtc cactctcctc
     2401 atcaatcagc cccagtatgc gtggctgaaa gagctggggc tccgcgagga aaacgagggc
     2461 gtgtataatg gaagctgggg aggccgggga gaggttatta cgacctattg ccctgctaac
     2521 aacgagccaa tagcaagagt ccgacaggcc agtgtggcag actatgaaga aactgtaaag
     2581 aaagcaagag aagcatggaa aatctgggca gatattcctg ctccaaaacg aggagaaata
     2641 gtaagacaga ttggcgatgc cttgcgggag aagatccaag tactaggaag cttggtgtct
     2701 ttggagatgg ggaaaatctt agtggaaggt gtgggtgaag ttcaggagta tgtggatatc
     2761 tgtgactatg ctgttggttt atcaaggatg attggaggac ctatcttgcc ttctgaaaga
     2821 tctggccatg cactgattga gcagtggaat cccgtaggcc tggttggaat catcacggca
     2881 ttcaatttcc ctgtggcagt gtatggttgg aacaacgcca tcgccatgat ctgtggaaat
     2941 gtctgcctct ggaaaggagc tccaaccact tccctcatta gtgtggctgt cacaaagata
     3001 atagccaagg ttctggagga caacaagctg cctggtgcaa tttgttcctt gacttgtggt
     3061 ggagcagata ttggcacagc aatggccaaa gatgaacgag tgaacctgct gtccttcact
     3121 gggagcactc aggtgggaaa acaggtgggc ctgatggtgc aggagaggtt tgggagaagt
     3181 ctgttggaac ttggaggaaa caatgccatt attgcctttg aagatgcaga cctcagctta
     3241 gttgttccat cagctctctt cgctgctgtg ggaacagctg gccagaggtg taccactgcg
     3301 aggcgactgt ttatacatga aagcatccat gatgaggttg taaacagact taaaaaggcc
     3361 tatgcacaga tccgagttgg gaacccatgg gaccctaatg ttctctatgg gccactccac
     3421 accaagcagg cagtgagcat gtttcttgga gcagtggaag aagcaaagaa agaaggtggc
     3481 acagtggtct atgggggcaa ggttatggat cgccctggaa attatgtaga accgacaatt
     3541 gtgacaggtc ttggccacga tgcgtccatt gcacacacag agacttttgc tccgattctc
     3601 tatgtcttta aattcaagaa tgaagaagag gtctttgcat ggaataatga agtaaaacag
     3661 ggactttcaa gtagcatctt taccaaagat ctgggcagaa tctttcgctg gcttggacct
     3721 aaaggatcag actgtggcat tgtaaatgtc aacattccaa caagtggggc tgagattgga
     3781 ggtgcctttg gaggagaaaa gcacactggt ggtggcaggg agtctggcag tgatgcctgg
     3841 aaacagtaca tgagaaggtc tacttgtact atcaactaca gtaaagacct tcctctggcc
     3901 caaggaatca agtttcaggg atccgactac aaagaccatg acggtgatta taaagatcat
     3961 gacatcgact acaaggatga cgatgacaag tagcggccgc gaaggatctg cgatcgctcc
     4021 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     4081 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     4141 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     4201 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg
     4261 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg
     4321 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta
     4381 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga
     4441 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg
     4501 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agatgaccga
     4561 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct
     4621 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat
     4681 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     4741 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     4801 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     4861 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     4921 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     4981 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     5041 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     5101 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagtcgacaa
     5161 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     5221 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     5281 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     5341 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     5401 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     5461 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     5521 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     5581 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     5641 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     5701 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa
     5761 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     5821 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc
     5881 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     5941 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     6001 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
     6061 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     6121 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     6181 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     6241 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc
     6301 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     6361 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg
     6421 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6481 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6541 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6601 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6661 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6721 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6781 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6841 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6901 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6961 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7021 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7081 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7141 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7201 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7261 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     7321 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7381 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7441 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7501 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7561 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7621 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7681 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7741 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7801 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7861 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7921 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7981 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8041 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8101 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8161 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8221 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8281 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8341 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8401 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8461 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8521 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8581 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     8641 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     8701 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     8761 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     8821 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     8881 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     8941 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     9001 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa
     9061 gctg
//
