LOCUS       V035993                 8368 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035993
VERSION     V035993
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    1798..1915
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        1960..2171
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             2184..2452
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             2495..2521
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2522..3775
                     /label="RAD52(NM_134424)"
                     /note="RAD52(NM_134424)"
                     /gene="RAD52"
     CDS             3791..3844
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             3851..4450
                     /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    4463..5051
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5125..5358
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5430..5551
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5570..5705
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(5743..5759)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5767..5783
                     /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(5791..5821)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5836..5857
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6145..6733)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6904..7764)
                     /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(7765..7869)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8343..8359
                     /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 ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca
     1981 cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc
     2041 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
     2101 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
     2161 ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta
     2221 cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct
     2281 cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc
     2341 cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc
     2401 ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt
     2461 gaccggcgcc tactctagag ctagcgccac catgtaccca tacgacgtac ctgattatgc
     2521 aatgtctggg actgaggaag caattcttgg aggacgtgac agccatcctg ctgctggcgg
     2581 cggctcagtg ttatgctttg gacagtgcca gtacacagca gaagagtacc aggccatcca
     2641 gaaggccctg aggcagaggc tgggcccaga atacataagt agccgcatgg ctggcggagg
     2701 ccagaaggtg tgctacattg agggtcatcg ggtaattaat ctggccaatg agatgtttgg
     2761 ttacaatggc tgggcacact ccatcacgca gcagaatgtg gattttgttg acctcaacaa
     2821 tggcaagttc tacgtgggag tctgtgcatt tgtgagggtc cagctgaagg atggttcata
     2881 tcatgaagat gttggttatg gtgttagtga gggcctcaag tccaaggctt tatctttgga
     2941 gaaggcaagg aaggaggcgg tgacagacgg gctgaagcga gccctcagga gttttgggaa
     3001 tgcacttgga aactgtattc tggacaaaga ctacctgaga tcactaaata agcttccacg
     3061 ccagttgcct cttgaagtgg atttaactaa agcgaagaga caagatcttg aaccgtctgt
     3121 ggaggaggca agatacaaca gctgccgacc gaacatggcc ctgggacacc cacagctgca
     3181 gcaggtgacc tccccttcca gacccagcca tgctgtgata ccggcggacc aggactgcag
     3241 ctcccgaagc ctgagctcat ccgccgtgga gagcgaggcc acgcaccagc ggaagctccg
     3301 gcagaagcag ctgcagcagc agttccggga gcggatggag aagcagcagg ttcgagtctc
     3361 cacgccgtca gctgagaaga gtgaggcagc gcctccggcc cctcctgtga cgcacagcac
     3421 tcctgtaact gtctcagaac cactcctgga gaaagacttc cttgcaggag tgactcaaga
     3481 attaatcaag actcttgaag acaactctga aaagtgggct gtgactcccg atgcagggga
     3541 tggtgtggtc aagccctcgt ctagagcaga cccagcccag acctctgaca cattagcctt
     3601 gaacaaccag atggtgaccc agaacaggac tccacacagc gtttgccacc agaaaccaca
     3661 agcaaaatct ggatcttggg acctccaaac ttatagcgct gaccaacgca caacaggaaa
     3721 ctgggaatct cataggaaga gccaggacat gaagaaaagg aaatatgatc catctggatc
     3781 cgcggccgct gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg
     3841 cccttccggg atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     3901 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     3961 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     4021 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     4081 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     4141 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     4201 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     4261 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     4321 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     4381 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     4441 cggtgcctga atctaggtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     4501 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     4561 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     4621 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     4681 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     4741 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4801 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     4861 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     4921 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4981 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     5041 cgcctccccg cctggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     5101 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga aaataagatc
     5161 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     5221 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     5281 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     5341 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     5401 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     5461 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5521 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     5581 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5641 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     5701 aggcctagac ttttgcagag acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt
     5761 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5821 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5881 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5941 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6001 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6061 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6121 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6181 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6241 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6301 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6361 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6421 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6481 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6541 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     6601 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6661 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6721 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6781 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6841 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6901 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6961 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7021 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7081 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7141 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7201 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7261 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7321 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7381 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7441 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7501 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7561 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7621 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7681 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7741 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7801 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7861 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     7921 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     7981 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     8041 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     8101 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     8161 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     8221 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     8281 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     8341 ttgtaaaacg acggccagtg ccaagctg
//
