LOCUS       V025762                 9105 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025762
VERSION     V025762
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..3959
                     /label="Atp5f1a(NM_023093)"
                     /note="Atp5f1a(NM_023093)"
                     /gene="Atp5f1a"
     CDS             3966..4031
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4075..4286
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4299..4567
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4594..5193
                     /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    5200..5788
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5862..6095
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6167..6288
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6307..6442
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6480..6496)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6504..6520
                     /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(6528..6558)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6573..6594
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6882..7470)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7641..8501)
                     /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(8502..8606)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9080..9096
                     /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 atgctgtccg tgcgcatcgc cgcggccgtg gcccgtgccc
     2341 tccctcgacg ggcgggactg gtctccaaaa atgccttggg gtcatctttt gttggtacaa
     2401 gaaatctcca tgcctctaac actcgacttc agaagactgg cactgccgag atgtcctcca
     2461 ttctcgagga gcggatcctt ggagctgata cttctgttga ccttgaagag actggccgtg
     2521 tcttaagcat tggtgatggt attgcccgag tgcatggact gaggaacgtt caagctgagg
     2581 agatggtaga gttttcttcc ggcttaaagg gtatgtccct gaacttggaa cccgacaatg
     2641 ttggagttgt cgtgtttggg aatgacaagc taattaaaga aggtgatatc gtgaagagaa
     2701 caggagccat cgtggacgtt ccagttggcg atgaactgtt gggccgtgta gttgatgccc
     2761 tgggaaatgc cattgatggg aagggtccag ttggttccaa gattcgcaga cgagtgggcc
     2821 tgaaagcccc tggaattatc ccccgaatct ctgtgcggga accaatgcag acgggcatca
     2881 aggctgtgga tagcctggtg ccgattggcc gaggtcagcg tgagctgatt attggagaca
     2941 gacagaccgg gaaaacctcg attgctattg acacaatcat caaccagaag cgtttcaatg
     3001 atgggactga cgagaagaag aaactgtact gcatctacgt tgctattggt cagaaacggt
     3061 ccaccgttgc tcagttggtg aagagactga cagacgcaga tgccatgaag tacaccattg
     3121 tggtgtcagc cactgcctct gatgctgccc cgcttcagta cttggctcct tactccggct
     3181 gctccatggg agagtatttc cgagataatg gcaagcacgc tctgatcatc tatgacgact
     3241 tatccaagca ggctgttgct taccgccaga tgtctctgct gctccgccga cccccgggtc
     3301 gagaggccta tcccggtgat gtgttttacc tacactctcg cctgctggag agagcagcca
     3361 agatgaacga ttcctttggt ggtggctctt tgactgcctt accagtcatt gaaacacagg
     3421 ctggtgatgt gtccgcctac attccaacaa acgttatttc catcaccgat ggacagatct
     3481 tcttggaaac agaattgttc tataaaggca tccgccctgc cattaatgtg ggcttgtctg
     3541 tgtcccgtgt cggatctgcc gcccagacca gagccatgaa gcaggtggca ggcaccatga
     3601 agctggagtt ggcccagtac cgggaggtcg ctgcttttgc ccagtttggt tctgatctgg
     3661 atgctgccac tcagcagctc ttgagccgtg gcgtgcgcct gaccgagctg ctaaagcaag
     3721 gacagtactc tcccatggct attgaagaac aggtggctgt catctatgca ggcgtccggg
     3781 gttatcttga taaactggag cccagtaaga tcacaaagtt cgagagtgct ttcttgtctc
     3841 atgttgtgag ccagcaccag agcctcttgg gcaatatcag gtctgatggg aaaatctcag
     3901 aacagtcgga tgcaaagctg aaggaaatcg taacaaactt cttggctggg tttgaacctg
     3961 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     4021 acgatgacaa gtagcggccg cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag
     4081 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg
     4141 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     4201 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     4261 gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac
     4321 gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc
     4381 ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga
     4441 gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc
     4501 tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt
     4561 acagatccaa gctgtgaccg gcgcctacgc tagatgaccg agtacaagcc cacggtgcgc
     4621 ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac
     4681 taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg
     4741 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     4801 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     4861 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     4921 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     4981 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     5041 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     5101 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     5161 acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca atcaacctct ggattacaaa
     5221 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     5281 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     5341 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     5401 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     5461 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     5521 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     5581 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     5641 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     5701 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     5761 cggatctccc tttgggccgc ctccccgcct ggtaccttta agaccaatga cttacaaggc
     5821 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     5881 ccaacgaaaa taagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
     5941 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     6001 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     6061 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
     6121 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
     6181 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6241 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
     6301 ctagctatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     6361 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     6421 tgaggaggct tttttggagg cctagacttt tgcagagacg gcccaaattc gtaatcatgg
     6481 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6541 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6601 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6661 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6721 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6781 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6841 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6901 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6961 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     7021 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     7081 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     7141 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     7201 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7261 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7321 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7381 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7441 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7501 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7561 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7621 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7681 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7741 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7801 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7861 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7921 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7981 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     8041 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     8101 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     8161 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     8221 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     8281 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     8341 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8401 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8461 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8521 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8581 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     8641 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     8701 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     8761 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     8821 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     8881 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     8941 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     9001 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     9061 ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctg
//
