LOCUS       V041893                 9718 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041893
VERSION     V041893
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..4572
                     /label="MINDY4(NM_032222)"
                     /note="MINDY4(NM_032222)"
                     /gene="MINDY4"
     CDS             4579..4644
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4688..4899
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4912..5180
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5207..5806
                     /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    5813..6401
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6475..6708
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6780..6901
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6920..7055
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7093..7109)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7117..7133
                     /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(7141..7171)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7186..7207
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7495..8083)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8254..9114)
                     /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(9115..9219)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9693..9709
                     /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 catggacagc ctcttcgtgg aggaggtggc cgcctccttg
     2341 gtcagggagt tcctcagcag aaagggctta aagaagacat gtgtgaccat ggaccaggaa
     2401 cgcccacgct ctgacctcag cataaacaac agaaatgatc ttcgaaaggt tttgcatctt
     2461 gaatttctct ataaggagaa caaggcaaag gaaaatcctc taaaaacaag ccttgaactc
     2521 atcaccagat actttctgga tcactttgga aatacggcta acaatttcac tcaagatacc
     2581 ccaatccctg cactctcagt tccaaagaaa aataacaaag tgccatcaag atgctcagag
     2641 actacactgg taaatatata tgacctttca gatgaagatg caggatggag aacatcattg
     2701 tcagaaacaa gcaaagccag acatgacaat cttgatggag atgtacttgg taattttgta
     2761 tcatctaaaa ggcccccgca caaaagtaag cccatgcaga cggtcccggg tgaaactcct
     2821 gtgttgactt ctgcatggga gaagatagac aagcttcact cggagccttc cttggatgtg
     2881 aagaggatgg gagagaattc caggccaaag tctggtctga ttgtgcgagg catgatgtct
     2941 gggcccatcg ccagctcccc acaggattct tttcacagac actatctgag acggtcctca
     3001 ccgtcaagca gctccaccca accccaagaa gagagccgga aggtccctga gctctttgtc
     3061 tgcacccaac aggacattct ggcttcgagc aacagctccc cctccaggac ctccctgggt
     3121 cagcttagtg aactgaccgt agaaaggcag aaaaccactg ccagcagccc tccccatctg
     3181 cccagcaaac ggctgccccc atgggacagg gccaggccga gggatccctc cgaggacacc
     3241 ccagcagtgg acggcagcac agacacggac aggatgccct tgaagctcta cttgcctggt
     3301 ggtaattcca ggatgaccca ggagaggctg gaaagagcgt tcaaacggca gggcagccag
     3361 cccgcacctg tcaggaaaaa tcagttgctg ccgtctgaca aggtggatgg tgagctgggt
     3421 gccctgcggc tcgaggatgt ggaggatgag ttgataaggg aagaggtcat cctgtcgcca
     3481 gtcccatcag tgctcaagtt gcagacagca tcaaaaccaa ttgacctctc agtagcaaag
     3541 gaaataaaga cccttctgtt tggttccagc ttttgctgtt tcaatgaaga atggaaactt
     3601 cagagttttt cctttagtaa cacagcctca ttaaaatacg gcatagtgca gaacaagggt
     3661 ggtccttgcg gagtcctggc agctgtccaa ggctgtgtcc tacagaaact cctgtttgaa
     3721 ggagatagca aagccgactg tgctcaggga ctgcagcctt cagatgccca ccggacccgc
     3781 tgcctcgtcc tggccctcgc agacattgtg tggcgggcag ggggccgaga gagagccgtt
     3841 gttgcactgg cttcgagaac acagcagttc agtccaacag ggaaatacaa agcagatgga
     3901 gtcttagaaa cacttacgct tcacagtttg acctgctatg aggacctggt gacttttctt
     3961 caacaaagca ttcatcagtt tgaagtgggc ccctatggct gcatcctgct caccctttct
     4021 gccatcctgt ccaggtctac agagctcatc cgccaggact ttgatgtccc caccagccac
     4081 ctgattggag cacatggcta ctgtacacag gaacttgtca atctgctcct gactgggaaa
     4141 gctgtgtcca acgttttcaa cgatgtggtt gagctggatt ctggggatgg gaacatcaca
     4201 cttctcagag gcattgctgc acgcagtgat attggcttct tatctctctt tgagcattac
     4261 aacatgtgcc aggttggctg cttcctgaag accccgaggt tccccatctg ggtggtttgc
     4321 agtgagagcc acttcagcat cctctttagc ctgcagccgg ggctcctgcg tgactggagg
     4381 actgagaggc tctttgactt gtactactac gatggcctgg ccaaccagca ggagcagatc
     4441 cggctgacca ttgacaccac ccaaaccatc tctgaggaca cagacaacga ccttgtccca
     4501 cccctcgagc tctgcatcag aaccaagtgg aagggggcat cagtgaactg gaacggctca
     4561 gaccccatcc tgggatccga ctacaaagac catgacggtg attataaaga tcatgacatc
     4621 gactacaagg atgacgatga caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc
     4681 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
     4741 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     4801 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     4861 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc
     4921 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc
     4981 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta
     5041 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc
     5101 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct
     5161 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagatga ccgagtacaa
     5221 gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc
     5281 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg
     5341 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     5401 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     5461 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     5521 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     5581 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     5641 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     5701 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     5761 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagtcg acaatcaacc
     5821 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     5881 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     5941 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     6001 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     6061 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     6121 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     6181 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     6241 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     6301 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     6361 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa
     6421 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     6481 ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg tctctctggt
     6541 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     6601 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     6661 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     6721 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
     6781 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     6841 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     6901 atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc attctccgcc
     6961 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     7021 attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag acggcccaaa
     7081 ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     7141 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     7201 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     7261 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     7321 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7381 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7441 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     7501 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     7561 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     7621 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     7681 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7741 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7801 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7861 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7921 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7981 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     8041 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     8101 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     8161 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     8221 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     8281 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     8341 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     8401 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     8461 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     8521 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     8581 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     8641 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     8701 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     8761 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     8821 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     8881 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     8941 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     9001 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     9061 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     9121 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     9181 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     9241 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     9301 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     9361 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     9421 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     9481 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     9541 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     9601 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     9661 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg
//
