LOCUS       V046461                 9628 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046461
VERSION     V046461
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..4482
                     /label="NDUFS1(NM_005006)"
                     /note="NDUFS1(NM_005006)"
                     /gene="NDUFS1"
     CDS             4489..4554
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4598..4809
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4822..5090
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5117..5716
                     /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    5723..6311
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6385..6618
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6690..6811
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6830..6965
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7003..7019)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7027..7043
                     /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(7051..7081)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7096..7117
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7405..7993)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8164..9024)
                     /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(9025..9129)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9603..9619
                     /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 catgttaagg atacctgtaa gaaaggcctt agtaggcctt
     2341 tctaagtctc ctaaaggatg tgttcgaaca actgccacag cagcaagcaa cttgattgaa
     2401 gtatttgttg atggtcagtc tgtcatggtg gaaccgggaa cgaccgtcct ccaagcttgt
     2461 gagaaggttg gcatgcagat ccctcgattc tgttatcatg aaaggttgtc tgttgctgga
     2521 aactgcagga tgtgccttgt tgaaattgag aaagccccta aggttgtagc tgcttgtgcc
     2581 atgccagtaa tgaagggttg gaatatccta acaaactcag aaaaatccaa aaaagccagg
     2641 gaaggtgtga tggagttctt attagcaaat cacccattgg actgtcctat ttgtgaccag
     2701 ggaggtgaat gtgatctgca ggaccagtcc atgatgtttg gaaatgatag gagccgattt
     2761 ttagagggga agcgtgctgt ggaagacaag aacattgggc cattggtaaa gaccatcatg
     2821 acaagatgta tacagtgtac tcgctgcatc aggtttgcaa gtgagattgc aggagtagat
     2881 gatttgggaa caacaggcag aggaaatgat atgcaagttg gcacatacat tgaaaagatg
     2941 ttcatgtctg aactgtctgg gaatatcatt gatatctgcc ctgtaggtgc cctaacctct
     3001 aagccctatg cctttactgc ccggccttgg gaaacaagaa agacagaatc cattgatgta
     3061 atggatgcgg ttggaagtaa tattgtggtt agcacaagaa ctggagaagt gatgaggatt
     3121 ttgccacgta tgcatgagga catcaatgaa gagtggatct ctgataaaac cagatttgcc
     3181 tatgatgggc taaaacgtca aagacttacc gagccaatgg tcagaaatga aaaagggctt
     3241 ttaacctata cttcttggga ggatgcgctc tctcgcgtag ctggaatgtt gcagagtttt
     3301 caaggcaaag atgtggcagc aattgcaggt ggcttggtgg atgctgaagc cctggtagct
     3361 ctcaaagatt tgcttaatag agtggactct gacaccttat gcactgaaga ggtcttcccc
     3421 actgcaggag ctggcacaga tttgcgttcc aattatcttc ttaatactac aattgctggt
     3481 gtggaagagg cagatgttgt tcttctggtt ggtacaaacc cacgttttga ggcaccactg
     3541 tttaatgcta gaattcgaaa gagctggctg cataatgact taaaagtggc ccttataggc
     3601 agtccagtgg acctcactta cacatatgac cacctgggag actcccccaa aattcttcaa
     3661 gacattgctt cgggaagcca tccatttagc caggtcctaa aggaagctaa aaaaccaatg
     3721 gtggttttag gcagttctgc actccaaaga aatgatggag cagcaattct tgcagctgtt
     3781 tctagcattg cacaaaagat tcggatgact agtggtgtta ctggtgattg gaaagttatg
     3841 aatatccttc ataggattgc aagtcaagta gctgctttgg accttggcta taagcctggg
     3901 gtggaagcaa ttcggaagaa ccctcccaag gtgctgtttc tcctgggagc agatggaggt
     3961 tgtatcacac gacaggattt gccaaaggat tgtttcatta tttatcaagg acatcatggt
     4021 gatgttgggg ctcccatagc tgatgttatt ctcccaggag ctgcttacac agagaagtct
     4081 gctacatatg tcaacactga gggtagagct cagcagacta aggtagcagt gacacctcct
     4141 ggcttggcaa gagaagactg gaaaattata agagcactct ctgagattgc tggaatgact
     4201 cttccatatg atactctgga tcaagtaagg aacagattgg aagaagtctc tcctaatctt
     4261 gttcgatatg atgatattga aggggctaat tacttccagc aagcaaatga gctctcaaag
     4321 ctagtgaacc agcagcttct tgctgaccca cttgttccac ctcagctaac tataaaagac
     4381 ttctacatga cagattcaat tagcagagcc tcacagacaa tggccaaatg tgtcaaagct
     4441 gtcacagagg gtgcccaggc agtagaggaa ccatccatat gcggatccga ctacaaagac
     4501 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtagcgg
     4561 ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca
     4621 gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc
     4681 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga
     4741 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc
     4801 agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc
     4861 tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc
     4921 tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg
     4981 gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt
     5041 gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga
     5101 ccggcgccta cgctagatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     5161 cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     5221 caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     5281 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     5341 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     5401 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     5461 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     5521 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     5581 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     5641 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     5701 caagcccggt gcctgagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     5761 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     5821 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     5881 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     5941 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     6001 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     6061 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     6121 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     6181 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     6241 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     6301 cgcctccccg cctggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     6361 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga aaataagatc
     6421 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     6481 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     6541 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     6601 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     6661 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     6721 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     6781 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     6841 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     6901 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     6961 aggcctagac ttttgcagag acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt
     7021 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     7081 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     7141 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     7201 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     7261 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     7321 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     7381 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     7441 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7501 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7561 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7621 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7681 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7741 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7801 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     7861 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7921 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7981 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     8041 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     8101 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     8161 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     8221 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     8281 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     8341 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     8401 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     8461 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     8521 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     8581 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8641 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8701 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8761 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8821 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8881 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8941 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     9001 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     9061 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     9121 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     9181 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     9241 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     9301 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     9361 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     9421 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     9481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     9541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     9601 ttgtaaaacg acggccagtg ccaagctg
//
