LOCUS       V020060                 9033 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020060
VERSION     V020060
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             2833..4143
                     /label="E2F2(NM_004091)"
                     /note="E2F2(NM_004091)"
                     /gene="E2F2"
     CDS             4159..4212
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             4222..4941
                     /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"
     misc_feature    4955..5543
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5750..6383
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6511..6527)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6535..6551
                     /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(6559..6589)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6604..6625
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6913..7501)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7672..8532)
                     /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(8533..8637)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8685..8819
                     /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 gtgaattcct cgagactagt
     2821 tctagagcca ccatgctgca agggccccgg gccttggctt cggccgctgg gcagaccccg
     2881 aaggtggtgc ccgcgatgag ccccacagag ctgtggccat ccggcctcag cagcccccag
     2941 ctctgcccag ctactgctac ctactacaca ccgctgtacc cgcagacggc gcctcccgca
     3001 gcggcgccag gcacctgcct cgacgccact ccccacggac ccgagggcca agttgtgcga
     3061 tgcctgccgg caggccggct gccggccaaa aggaagctgg atctggaggg gattgggagg
     3121 cccgtcgtcc ctgagttccc aacccccaag gggaagtgca tcagagtgga tggcctcccc
     3181 agccccaaaa cccccaaatc ccccggggag aagactcggt atgacacttc gctggggctg
     3241 ctcaccaaga agttcattta cctcctgagc gagtcagagg atggggtcct ggacctgaac
     3301 tgggccgctg aggtgctgga cgtgcagaag cggcgcatct atgacatcac caacgtgctg
     3361 gaaggcatcc agctcatccg caagaaggcc aagaacaaca tccagtgggt aggcagggga
     3421 atgtttgaag accccaccag acctgggaag cagcaacagc tggggcagga gctgaaggag
     3481 ctgatgaaca cggagcaggc cttggaccag ctcatccaga gctgctctct gagcttcaag
     3541 cacctgactg aggacaaggc caacaagagg ctggcctatg tgacttacca ggatatccgt
     3601 gctgttggca actttaagga gcagacagtg attgccgtca aggcccctcc gcagacgaga
     3661 ctggaagtgc ccgacaggac tgaggacaac ctgcagatat atctcaagag cacccaaggg
     3721 cccatcgaag tctacctgtg cccagaggag gtgcaggagc cggacagtcc ttccgaggag
     3781 cctctcccct ctacctccac cctctgcccc agccctgact ctgcccagcc cagcagcagc
     3841 accgacccta gcatcatgga gcccacagca tcctcagtgc cagcaccagc gccaaccccc
     3901 cagcaggccc caccgcctcc atccctggtc cccttggagg ctactgacag cctgctggag
     3961 ctgccgcacc cactcctgca gcagactgag gaccagttcc tgtccccgac cctggcgtgc
     4021 agctcccctc tgatcagctt ctccccatcc ttggaccagg acgactacct gtggggcttg
     4081 gaggcgggtg agggcatcag cgatctcttc gactcctacg accttgggga cctgttgatt
     4141 aatggatccg cggccgctga gggcagagga agtcttctaa catgcggtga cgtggaggag
     4201 aatcccggcc ctaccggtat catggtgagc aagggcgagg agctgttcac cggggtggtg
     4261 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     4321 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     4381 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     4441 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     4501 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     4561 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     4621 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     4681 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     4741 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     4801 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     4861 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     4921 atggacgagc tgtacaagta aacgcgtctg gaacaatcaa cctctggatt acaaaatttg
     4981 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     5041 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     5101 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     5161 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     5221 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     5281 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     5341 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     5401 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     5461 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     5521 ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct agaaaaacat
     5581 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca
     5641 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact
     5701 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact ggaagggcta
     5761 attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac
     5821 ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact gacctttgga
     5881 tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa taaaggagag
     5941 aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagtg
     6001 ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga gctgcatccg
     6061 gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc tggggacttt
     6121 ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa
     6181 gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct
     6241 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     6301 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     6361 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     6421 acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc gtttaccgtc
     6481 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6541 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6601 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6661 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6721 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6781 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6841 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6901 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6961 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7021 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7081 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7141 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7201 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7261 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7321 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     7381 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7441 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7501 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7561 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7621 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7681 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7741 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7801 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7861 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7921 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7981 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     8041 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8101 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8161 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8221 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8281 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8341 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8401 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8461 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8521 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8581 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8641 atttccccga aaagtgccac ctgacgtcga cggatcggga gatcaacttg tttattgcag
     8701 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     8761 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca
     8821 actggataac tcaagctaac caaaatcatc ccaaacttcc caccccatac cctattacca
     8881 ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa ttattttcat
     8941 tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagttttt aaagaaattg
     9001 tatttgttaa atatgtacta caaacttagt agt
//
