LOCUS       V024220                 9439 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024220
VERSION     V024220
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        9..235
                     /note="Rous sarcoma virus enhancer/promoter"
                     /label="RSV promoter"
     LTR             236..416
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    463..588
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1081..1314
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1499..1543
                     /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    1810..1926
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2130..3312
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     promoter        3329..3347
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             3369..4262
                     /label="AZGP1(NM_001185)"
                     /note="AZGP1(NM_001185)"
                     /gene="AZGP1"
     misc_feature    4271..4844
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES"
     CDS             4878..5276
                     /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    5291..5879
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5966..6199
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6271..6392
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6432..6567
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(6588..6606)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(6616..6632)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      6774..7229
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        7255..7359
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             7360..8220
                     /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"
     rep_origin      8391..8979
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    9267..9288
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        9303..9333
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    9341..9357
                     /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"
     primer_bind     9365..9381
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        9402..9420
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /label="T3 promoter"
ORIGIN
        1 aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta
       61 gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg
      121 gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac
      181 tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc
      241 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
      301 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
      361 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
      421 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
      481 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
      541 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
      601 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
      661 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
      721 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
      781 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
      841 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
      901 aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat
      961 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     1021 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     1081 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     1141 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     1201 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     1261 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     1321 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     1381 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     1441 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     1501 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     1561 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     1621 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     1681 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
     1741 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta
     1801 tcggttaact tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     1861 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaattcaa
     1921 aattttatcg atgcctcccc gtcaccaccc cccccaaccc gccccgaccg gagctgagag
     1981 taattcatac aaaaggactc gcccctgcct tggggaatcc cagggaccgt cgttaaactc
     2041 ccactaacgt agaacccaga gatcgctgcg ttcccgcccc ctcacccgcc cgctctcgtc
     2101 atcactgagg tggagaagag catgcgtgag gctccggtgc ccgtcagtgg gcagagcgca
     2161 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga
     2221 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     2281 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     2341 ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta
     2401 cgggttatgg cccttgcgtg ccttgaatta cttccacgcc cctggctgca gtacgtgatt
     2461 cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga
     2521 gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga
     2581 atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat
     2641 ttttgatgat atcctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa
     2701 gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc
     2761 cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg
     2821 tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg
     2881 ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt
     2941 cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt
     3001 gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc
     3061 acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg
     3121 tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag
     3181 actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag
     3241 tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt
     3301 caggtgtcgt gaaaactacc cctctagata atacgactca ctatagggcc cgggttggat
     3361 ccgccaccat ggtaagaatg gtgcctgtcc tgctgtctct gctgctgctt ctgggtcctg
     3421 ctgtccccca ggagaaccaa gatggtcgtt actctctgac ctatatctac actgggctgt
     3481 ccaagcatgt tgaagacgtc cccgcgtttc aggcccttgg ctcactcaat gacctccagt
     3541 tctttagata caacagtaaa gacaggaagt ctcagcccat gggactctgg agacaggtgg
     3601 aaggaatgga ggattggaag caggacagcc aacttcagaa ggccagggag gacatcttta
     3661 tggagaccct gaaagacatc gtggagtatt acaacgacag taacgggtct cacgtattgc
     3721 agggaaggtt tggttgtgag atcgagaata acagaagcag cggagcattc tggaaatatt
     3781 actatgatgg aaaggactac attgaattca acaaagaaat cccagcctgg gtccccttcg
     3841 acccagcagc ccagataacc aagcagaagt gggaggcaga accagtctac gtgcagcggg
     3901 ccaaggctta cctggaggag gagtgccctg cgactctgcg gaaatacctg aaatacagca
     3961 aaaatatcct ggaccggcaa gatcctccct ctgtggtggt caccagccac caggccccag
     4021 gagaaaagaa gaaactgaag tgcctggcct acgacttcta cccagggaaa attgatgtgc
     4081 actggactcg ggccggcgag gtgcaggagc ctgagttacg gggagatgtt cttcacaatg
     4141 gaaatggcac ttaccagtcc tgggtggtgg tggcagtgcc cccgcaggac acagccccct
     4201 actcctgcca cgtgcagcac agcagcctgg cccagcccct cgtggtgccc tgggaggcca
     4261 gctaggctag cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
     4321 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     4381 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     4441 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     4501 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     4561 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     4621 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     4681 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     4741 ggcctcggta cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     4801 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacactagag
     4861 atccaccggt cgccaccatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa
     4921 gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag
     4981 ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac
     5041 cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt
     5101 gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac
     5161 aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc
     5221 cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaagcgg
     5281 ccgcgtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     5341 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     5401 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     5461 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     5521 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     5581 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     5641 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     5701 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     5761 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     5821 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     5881 tggaattcga gctcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     5941 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     6001 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     6061 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     6121 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     6181 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     6241 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     6301 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     6361 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     6421 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     6481 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     6541 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc
     6601 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     6661 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     6721 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     6781 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     6841 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     6901 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     6961 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     7021 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     7081 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     7141 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     7201 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     7261 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     7321 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     7381 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     7441 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     7501 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     7561 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     7621 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     7681 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     7741 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     7801 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     7861 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     7921 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     7981 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     8041 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     8101 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     8161 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     8221 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     8281 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     8341 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     8401 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     8461 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     8521 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     8581 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     8641 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     8701 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     8761 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     8821 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     8881 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     8941 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     9001 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     9061 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     9121 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     9181 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     9241 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     9301 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     9361 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
     9421 gaacaaaagc tggagctgc
//
