LOCUS       V044759                 9548 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044759
VERSION     V044759
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             2290..3690
                     /label="Smad2(NM_010754)"
                     /note="Smad2(NM_010754)"
                     /gene="Smad2"
     CDS             3727..4443
                     /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             4450..4467
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     promoter        4518..4729
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4742..5010
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5037..5636
                     /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    5643..6231
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6305..6538
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6610..6731
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6750..6885
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6923..6939)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6947..6963
                     /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(6971..7001)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7016..7037
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7325..7913)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8084..8944)
                     /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(8945..9049)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9523..9539
                     /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 agagccacca tgtcgtccat cttgccattc actccgccag tggtgaagag acttctggga
     2341 tggaaaaaat cagccggtgg gtctggagga gcaggtggtg gagagcagaa tggacaggaa
     2401 gaaaagtggt gtgaaaaagc agtgaaaagt ctggtgaaaa agctaaagaa aacaggacgg
     2461 ttagatgagc ttgagaaagc catcaccact cagaattgca atactaaatg tgtcaccata
     2521 ccaagcactt gctctgaaat ttggggactg agtacagcaa atacggtaga tcagtgggac
     2581 acaacaggcc tttacagctt ctctgaacaa accaggtctc ttgatggccg tcttcaggtt
     2641 tcacaccgga aagggttgcc acatgttata tattgccggc tctggcgctg gccggacctt
     2701 cacagtcatc atgagctcaa ggcaatcgaa aactgcgaat atgcttttaa tctgaaaaaa
     2761 gatgaagtgt gtgtaaatcc gtaccactac cagagagttg agaccccagt cttgcctcca
     2821 gtcttagtgc ctcggcacac ggagattcta acagaactgc cgcccctgga tgactacacc
     2881 cactccattc cagaaaacac aaatttccca gcaggaattg agccacagag taattacatc
     2941 ccagaaacac caccacctgg atatatcagt gaagatggag aaacaagtga ccaacagttg
     3001 aaccaaagta tggacacagg ctctccggct gaactgtctc ctactactct ctctcctgtt
     3061 aatcacagct tggatttgca gccagttact tactcggaac ctgcattctg gtgttcaatc
     3121 gcatactatg aactaaacca gagggttgga gagaccttcc atgcgtcaca gccctcgctc
     3181 actgtagacg gcttcacaga cccatcaaac tcggagaggt tctgcttagg cttgctctcc
     3241 aacgttaacc gaaatgccac tgtagaaatg acaagaagac atataggaag gggagtgcgc
     3301 ttgtattaca taggtgggga agtgtttgct gagtgcctaa gtgatagtgc aatctttgtg
     3361 cagagcccca actgtaacca gagatacggc tggcaccctg caacagtgtg taagatccca
     3421 ccaggctgta acctgaagat cttcaacaac caagaatttg ctgctcttct ggctcagtct
     3481 gtcaaccagg gttttgaagc cgtttatcag ctaacccgaa tgtgcaccat aagaatgagt
     3541 tttgtgaagg gctggggagc agaatatcgg aggcagacag taacaagtac tccttgctgg
     3601 attgaacttc atctgaatgg ccctctgcag tggctggaca aagtattaac tcagatggga
     3661 tccccttcag tgcgatgctc aagcatgtcg gaattcggag gtggcggaag cggcggtgga
     3721 ggtagtatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     3781 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     3841 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     3901 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     3961 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     4021 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     4081 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     4141 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     4201 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     4261 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     4321 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     4381 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     4441 aagctcgagc accaccacca ccaccactga ggatccgcgg ccgcgaagga tctgcgatcg
     4501 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
     4561 aggggtcggc aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
     4621 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
     4681 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg
     4741 aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg
     4801 gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta
     4861 ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc
     4921 tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt
     4981 ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagatga
     5041 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca
     5101 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc
     5161 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     5221 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     5281 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     5341 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     5401 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     5461 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     5521 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     5581 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagtcg
     5641 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     5701 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     5761 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     5821 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     5881 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     5941 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     6001 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     6061 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     6121 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     6181 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct
     6241 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     6301 ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg
     6361 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     6421 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     6481 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     6541 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     6601 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6661 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6721 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc
     6781 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     6841 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag
     6901 acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6961 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     7021 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     7081 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     7141 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     7201 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     7261 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     7321 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     7381 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     7441 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     7501 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     7561 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     7621 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     7681 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7741 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     7801 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7861 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7921 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7981 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     8041 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     8101 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     8161 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     8221 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     8281 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     8341 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     8401 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     8461 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     8521 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     8581 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     8641 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     8701 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8761 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8821 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8881 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8941 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     9001 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     9061 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     9121 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     9181 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     9241 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     9301 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     9361 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     9421 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     9481 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     9541 ccaagctg
//
