LOCUS       V021278                 9132 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021278
VERSION     V021278
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             2815..3783
                     /label="CD47(NM_001777)"
                     /note="CD47(NM_001777)"
                     /gene="CD47"
     misc_feature    3803..4375
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES"
     CDS             4377..5087
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK*"
                     /label="mCherry"
     misc_feature    5101..5689
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5896..6529
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6657..6673)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6681..6697
                     /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(6705..6735)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6750..6771
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7059..7647)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7818..8678)
                     /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(8679..8783)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8831..8965
                     /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 tctatttccg gtgaattcgc caccatgtgg
     2821 cccctggtag cggcgctgtt gctgggctcg gcgtgctgcg gatcagctca gctactattt
     2881 aataaaacaa aatctgtaga attcacgttt tgtaatgaca ctgtcgtcat tccatgcttt
     2941 gttactaata tggaggcaca aaacactact gaagtatacg taaagtggaa atttaaagga
     3001 agagatattt acacctttga tggagctcta aacaagtcca ctgtccccac tgactttagt
     3061 agtgcaaaaa ttgaagtctc acaattacta aaaggagatg cctctttgaa gatggataag
     3121 agtgatgctg tctcacacac aggaaactac acttgtgaag taacagaatt aaccagagaa
     3181 ggtgaaacga tcatcgagct aaaatatcgt gttgtttcat ggttttctcc aaatgaaaat
     3241 attcttattg ttattttccc aatttttgct atactcctgt tctggggaca gtttggtatt
     3301 aaaacactta aatatagatc cggtggtatg gatgagaaaa caattgcttt acttgttgct
     3361 ggactagtga tcactgtcat tgtcattgtt ggagccattc ttttcgtccc aggtgaatat
     3421 tcattaaaga atgctactgg ccttggttta attgtgactt ctacagggat attaatatta
     3481 cttcactact atgtgtttag tacagcgatt ggattaacct ccttcgtcat tgccatattg
     3541 gttattcagg tgatagccta tatcctcgct gtggttggac tgagtctctg tattgcggcg
     3601 tgtataccaa tgcatggccc tcttctgatt tcaggtttga gtatcttagc tctagcacaa
     3661 ttacttggac tagtttatat gaaatttgtg gcttccaatc agaagactat acaacctcct
     3721 aggaaagctg tagaggaacc ccttaatgca ttcaaagaat caaaaggaat gatgaatgat
     3781 gaataagcgg ccgcggatcc cgcccctctc cctccccccc ccctaacgtt actggccgaa
     3841 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     3901 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     3961 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     4021 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     4081 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     4141 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     4201 tcacctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     4261 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaacg
     4321 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg
     4381 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     4441 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     4501 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     4561 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     4621 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     4681 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     4741 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     4801 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     4861 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     4921 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     4981 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     5041 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtgaacg cgtctggaac
     5101 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     5161 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     5221 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     5281 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     5341 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     5401 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     5461 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     5521 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     5581 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     5641 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattaat
     5701 tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca gcagctacca
     5761 atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt ccagtcacac
     5821 ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     5881 aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat atccttgatc
     5941 tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca ccagggccag
     6001 gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt gagccagata
     6061 aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg agcctgcatg
     6121 ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc ctagcatttc
     6181 atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat cgagcttgct
     6241 acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg actggggagt
     6301 ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg gtctctctgg
     6361 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     6421 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     6481 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
     6541 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     6601 ccttgacatt gctagcgttt accgtcgacc tctagctaga gcttggcgta atcatggtca
     6661 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     6721 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     6781 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6841 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6901 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6961 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     7021 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7081 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7141 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7201 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7261 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7321 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7381 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7441 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     7501 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7561 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7621 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7681 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7741 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7801 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7861 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7921 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7981 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8041 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8101 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8161 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8221 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8281 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8341 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8401 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8461 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8521 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8581 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     8641 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8701 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8761 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     8821 tcgggagatc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     8881 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     8941 caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa
     9001 acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac
     9061 ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac
     9121 aaacttagta gt
//
