LOCUS       V029426                 9301 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029426
VERSION     V029426
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /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    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2843..3997
                     /label="Dlk1(NM_010052)"
                     /note="Dlk1(NM_010052)"
                     /gene="Dlk1"
     CDS             4004..4069
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4089..4588
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4609..5208
                     /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    5222..5810
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6017..6650
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6779..6795)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6803..6819
                     /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(6827..6857)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6872..6893
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7181..7769)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7940..8800)
                     /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(8801..8905)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8953..9087
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag
     2821 ctcaagcttc gaattcgcca ccatgatcgc gaccggagcc ctcctgcgcg tcctcttgct
     2881 cctgctggct ttcggccaca gcacctatgg ggctgaatgc gacccaccct gtgaccccca
     2941 gtatggattc tgcgaggctg acaatgtctg caggtgccat gttggctggg agggtcccct
     3001 ctgtgacaag tgtgtaactg cccctggctg tgtcaatgga gtctgcaagg aaccatggca
     3061 gtgcatctgc aaggatggct gggacgggaa attctgcgaa atagacgttc gggcttgcac
     3121 ctcaaccccc tgcgccaaca atggaacttg cgtggacctg gagaaaggcc agtacgaatg
     3181 ctcctgcaca cctgggttct ctggaaagga ctgccagcac aaggctgggc cctgcgtgat
     3241 caatggttct ccctgccagc acggaggcgc ctgcgtggat gatgagggcc aggcctcgca
     3301 tgcttcctgc ctgtgccccc ctggcttctc aggcaacttc tgtgagatcg tagccgcaac
     3361 caacagctgt acccctaacc catgcgagaa cgatggcgtc tgcaccgaca tcgggggtga
     3421 cttccgttgc cgctgcccag ctggattcgt cgacaagacc tgcagccgcc cggtgagcaa
     3481 ctgcgccagt ggcccgtgcc agaacggggg cacctgcctc cagcacaccc aggtgagctt
     3541 cgagtgtctg tgcaagcccc cgttcatggg tcccacgtgc gcgaagaagc gcggggctag
     3601 ccccgtgcag gtcacccacc tgcccagcgg ctatgggctc acctaccgcc tgacccccgg
     3661 ggtgcacgag ctgcctgttc agcagcccga gcaacacatc ctgaaggtgt ccatgaaaga
     3721 gctcaacaag agtacccctc tcctcaccga gggacaggcc atctgcttca ccatcctggg
     3781 cgtgctcacc agcctggtgg tgctgggcac cgtggccatc gtctttctca acaagtgcga
     3841 aacctgggtg tccaacctgc gctacaacca catgcttcgc aagaagaaga acctcctgtt
     3901 gcagtataac agcggcgagg agctggcggt caatatcatc ttccccgaga agattgacat
     3961 gaccactttc aacaaggagg ctggtgatga ggagatcgga tccgactaca aagaccatga
     4021 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt aatctagata
     4081 attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc
     4141 cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac
     4201 cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc
     4261 cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga
     4321 agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt
     4381 aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct
     4441 gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg
     4501 aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt
     4561 ctcctcttcc tcatctccgg gcctttcgac ctgcagccca agcttaccat gaccgagtac
     4621 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     4681 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     4741 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     4801 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     4861 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     4921 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     4981 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     5041 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     5101 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     5161 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacc gcgtctggaa
     5221 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     5281 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     5341 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     5401 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     5461 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     5521 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     5581 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     5641 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     5701 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     5761 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattaa
     5821 ttctgcagtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
     5881 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca
     5941 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     6001 aaagaaaaga ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat
     6061 ctgtggatct accacacaca aggctacttc cctgattagc agaactacac accagggcca
     6121 ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagccagat
     6181 aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt gagcctgcat
     6241 gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg cctagcattt
     6301 catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata tcgagcttgc
     6361 tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg gactggggag
     6421 tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg ggtctctctg
     6481 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     6541 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     6601 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
     6661 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     6721 gccttgacat tgctagcgtt ttaccgtcga cctctagcta gagcttggcg taatcatggt
     6781 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6841 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6901 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6961 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     7021 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     7081 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     7141 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7201 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7261 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7321 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7381 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7441 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7501 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7561 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7621 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7681 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7741 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7801 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7861 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7921 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7981 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     8041 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     8101 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     8161 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8221 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     8281 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     8341 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8401 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8461 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8521 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8581 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8641 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8701 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8761 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8821 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8881 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     8941 gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     9001 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     9061 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc
     9121 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa
     9181 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact
     9241 acaaacttag tagtttttaa agaaattgta tttgttaaat atgtactaca aacttagtag
     9301 t
//
