LOCUS       V044091                 9025 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044091
VERSION     V044091
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             2302..4074
                     /label="GBP2(NM_004120)"
                     /note="GBP2(NM_004120)"
                     /gene="GBP2"
     CDS             4081..4146
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4190..4401
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4414..4682
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4715..5113
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     misc_feature    5120..5708
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5782..6015
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6087..6208
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6227..6362
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6400..6416)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6424..6440
                     /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(6448..6478)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6493..6514
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6802..7390)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7561..8421)
                     /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(8422..8526)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9000..9016
                     /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 agagctagcg aattcgccac catggctcca gagatcaact tgccgggccc aatgagcctc
     2341 attgataaca ctaaagggca gctggtggtg aatccagaag ctctgaagat cctatctgca
     2401 attacgcagc ctgtggtggt ggtggcgatt gtgggcctct atcgcacagg caaatcctac
     2461 ctgatgaaca agctggctgg gaagaaaaac ggcttctctc taggctccac agtgaagtct
     2521 cacaccaagg gaatctggat gtggtgtgtg cctcatccca agaagccaga acacacccta
     2581 gttctgctcg acactgaggg cctgggagat atagagaagg gtgacaatga gaatgactcc
     2641 tggatctttg ccttggccat cctcctgagc agcaccttcg tgtacaatag catgggaacc
     2701 atcaaccagc aggccatgga ccaacttcac tatgtgacag agctgacaga tcgaatcaag
     2761 gcaaactcct cacctggtaa caattctgta gacgactcag ctgactttgt gagctttttt
     2821 ccagcatttg tgtggactct cagagatttc accctggaac tggaagtaga tggagaaccc
     2881 atcactgctg atgactactt ggagctttcg ctaaagctaa gaaaaggtac tgataagaaa
     2941 agtaaaagct ttaatgatcc tcggttgtgc atccgaaagt tcttccccaa gaggaagtgc
     3001 ttcgtcttcg attggcccgc tcctaagaag taccttgctc acctagagca gctaaaggag
     3061 gaagagctga accctgattt catagaacaa gttgcagaat tttgttccta catcctcagc
     3121 cattccaatg tcaagactct ttcaggtggc attccagtca atgggcctcg tctagagagc
     3181 ctggtgctga cctacgtcaa tgccatcagc agtggggatc taccctgcat ggagaacgca
     3241 gtcctggcct tggcccagat agagaactca gccgcagtgg aaaaggctat tgcccactat
     3301 gaacagcaga tgggccagaa ggtgcagctg cccacggaaa ccctccagga gctgctggac
     3361 ctgcacaggg acagtgagag agaggccatt gaagtcttca tgaagaactc tttcaaggat
     3421 gtggaccaaa tgttccagag gaaattaggg gcccagttgg aagcaaggcg agatgacttt
     3481 tgtaagcaga attccaaagc atcatcagat tgttgcatgg ctttacttca ggatatattt
     3541 ggccctttag aagaagatgt caagcaggga acattttcta aaccaggagg ttaccgtctc
     3601 tttactcaga agctgcagga gctgaagaat aagtactacc aggtgccaag gaaggggata
     3661 caggccaaag aggtgctgaa aaaatatttg gagtccaagg aggatgtggc tgatgcactt
     3721 ctacagactg atcagtcact ctcagaaaag gaaaaagcga ttgaagtgga acgtataaag
     3781 gctgaatctg cagaagctgc aaagaaaatg ttggaggaaa tacaaaagaa gaatgaggag
     3841 atgatggaac agaaagagaa gagttatcag gaacatgtga aacaattgac tgagaagatg
     3901 gagagggaca gggcccagtt aatggcagag caagagaaga ccctcgctct taaacttcag
     3961 gaacaggaac gccttctcaa ggagggattc gagaatgaga gcaagagact tcaaaaagac
     4021 atatgggata tccagatgag aagcaaatca ttggagccaa tatgtaacat actcggatcc
     4081 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat
     4141 gacaagtagc ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc
     4201 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag
     4261 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     4321 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
     4381 gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc
     4441 cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc
     4501 tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg
     4561 ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc
     4621 ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga
     4681 tccaagctgt gaccggcgcc tacgctaggc caccatggcc aagcctttgt ctcaagaaga
     4741 atccaccctc attgaaagag caacggctac aatcaacagc atccccatct ctgaagacta
     4801 cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg tcaatgtata
     4861 tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc
     4921 agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca tcttgagccc
     4981 ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag ccatagtgaa
     5041 ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct ctggttatgt
     5101 gtgggagggc taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     5161 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     5221 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     5281 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     5341 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     5401 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     5461 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     5521 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     5581 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     5641 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     5701 ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     5761 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaaa taagatctgc
     5821 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     5881 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     5941 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     6001 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa
     6061 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat
     6121 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     6181 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac
     6241 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     6301 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     6361 cctagacttt tgcagagacg gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg
     6421 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6481 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6541 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6601 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6661 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6721 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6781 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6841 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6901 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6961 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     7021 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7081 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7141 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7201 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     7261 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7321 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7381 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7441 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7501 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7561 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7621 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7681 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7741 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7801 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7861 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7921 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7981 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8041 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8101 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8161 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8221 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8281 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8341 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8401 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8461 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8521 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     8581 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     8641 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     8701 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     8761 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     8821 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     8881 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     8941 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     9001 taaaacgacg gccagtgcca agctg
//
