LOCUS       V028752                 9785 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028752
VERSION     V028752
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             2291..3007
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             3023..4639
                     /label="TRIM26(NM_003449)"
                     /note="TRIM26(NM_003449)"
                     /gene="TRIM26"
     CDS             4646..4711
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4755..4966
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4979..5247
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5274..5873
                     /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    5880..6468
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6542..6775
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6847..6968
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6987..7122
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7160..7176)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7184..7200
                     /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(7208..7238)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7253..7274
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7562..8150)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8321..9181)
                     /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(9182..9286)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9760..9776
                     /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 gaacgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     2341 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     2401 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     2461 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     2521 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     2581 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2641 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2701 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     2761 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     2821 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2881 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     2941 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     3001 gtacaagggc ggtggaggca gtatggccac gtcagcccca ctacggagcc tggaagagga
     3061 ggtgacctgc tccatctgtc ttgattacct gcgggaccct gtgaccattg actgtggcca
     3121 cgtcttctgc cgcagctgca ccacagacgt ccgccccatc tcagggagcc gccccgtctg
     3181 cccactctgc aagaagcctt ttaagaagga gaacatccga cccgtgtggc aactggccag
     3241 cctggtggag aacattgagc ggctgaaggt ggacaagggc aggcagccgg gagaggtgac
     3301 ccgggagcag caggatgcaa agttgtgcga gcgacaccga gagaagctgc actactactg
     3361 tgaggacgac gggaagctgc tgtgcgtgat gtgccgggag tcccgggagc acaggcccca
     3421 cacggccgtc ctcatggaga aggccgccca gccccacagg gaaaaaatcc tgaaccacct
     3481 gagtacccta aggagggaca gagacaaaat tcagggcttc caggcaaagg gagaagctga
     3541 tatcctggcc gcgctgaaga agctccagga ccagaggcag tacattgtgg ctgagtttga
     3601 gcagggtcat cagttcctga gggagcggga ggaacacctg ctggaacagc tggcgaagct
     3661 ggagcaggag ctcacggagg gcagggagaa gttcaagagc cggggcgtcg gggagcttgc
     3721 ccggctggcc ctggtcatct ccgaactgga gggcaaggcg cagcagccag ctgcagagct
     3781 catgcaggac acgagagact tcctaaacag gtatccacgg aagaagttct gggttgggaa
     3841 acccattgct cgagtggtta aaaaaaagac cggagaattc tcagataaac tcctctctct
     3901 gcaacgaggc ctgagggaat tccaggggaa gctgctgaga gacttggaat ataagacagt
     3961 gagcgtcacc ctggacccac agtcggccag tgggtacctg cagctgtcag aggactggaa
     4021 gtgcgtgacc tacaccagcc tgtacaagag tgcctacctg cacccccagc agtttgactg
     4081 tgagcctggg gtgctaggca gcaagggctt cacctggggc aaggtctact gggaagtgga
     4141 agtggagagg gagggctggt ctgaggatga agaagagggg gatgaggagg aagagggaga
     4201 agaggaggag gaggaagagg aggccggcta tggggatgga tatgacgact gggaaacgga
     4261 cgaagatgag gaatcgttgg gcgatgaaga ggaagaagag gaggaggaag aggaggaagt
     4321 tctggaaagc tgcatggtgg gggtggctag agactctgtg aagaggaagg gagacctctc
     4381 cctgcggcca gaggatggcg tgtgggcgct gcgcctctcc tcctccggca tctgggccaa
     4441 caccagcccc gaggctgagc ttttcccagc actgcggccc cggagagtgg gcatcgccct
     4501 ggattatgaa gggggcaccg tgactttcac caacgcagag tcacaggaac tcatctacac
     4561 cttcactgcc accttcaccc ggcgcctggt ccccttcctg tggctcaagt ggccaggaac
     4621 acgcctcctg ctaagacccg gatccgacta caaagaccat gacggtgatt ataaagatca
     4681 tgacatcgac tacaaggatg acgatgacaa gtagcggccg cgaaggatct gcgatcgctc
     4741 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     4801 ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     4861 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     4921 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg
     4981 ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt
     5041 gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt
     5101 aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag
     5161 actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc
     5221 gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagatgaccg
     5281 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc
     5341 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca
     5401 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     5461 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     5521 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     5581 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     5641 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     5701 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     5761 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     5821 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca
     5881 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     5941 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     6001 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     6061 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     6121 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     6181 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     6241 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     6301 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     6361 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     6421 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta
     6481 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     6541 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct
     6601 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     6661 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     6721 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     6781 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     6841 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     6901 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     6961 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt
     7021 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     7081 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg
     7141 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     7201 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     7261 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     7321 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     7381 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     7441 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     7501 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     7561 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     7621 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7681 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7741 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7801 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7861 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7921 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7981 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     8041 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     8101 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     8161 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     8221 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     8281 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     8341 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     8401 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     8461 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     8521 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     8581 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     8641 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     8701 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8761 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8821 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8881 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8941 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     9001 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     9061 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     9121 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     9181 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     9241 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     9301 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     9361 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     9421 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     9481 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     9541 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     9601 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     9661 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     9721 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca
     9781 agctg
//
