LOCUS       V029521                 9365 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029521
VERSION     V029521
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    1798..1915
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2005..2208
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2297..4117
                     /label="RPN1(NM_002950)"
                     /note="RPN1(NM_002950)"
                     /gene="RPN1"
     promoter        4168..4379
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4392..4660
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4689..4698
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4719..5381
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     misc_feature    5460..6048
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6122..6355
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6427..6548
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6567..6702
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6740..6756)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6764..6780
                     /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(6788..6818)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6833..6854
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7142..7730)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7901..8761)
                     /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(8762..8866)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9340..9356
                     /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 ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagcgaattc gccaccatgg aggcgccagc cgccggcttg tttctgctcc tgttgcttgg
     2341 gacttgggcc ccggcgccgg gcagcgcctc ctccgaggca ccgccgctga tcaatgagga
     2401 cgtgaagcgc acagtggacc taagcagcca cctggctaag gtgacggccg aggtggtcct
     2461 ggcgcacctg ggcggcggct ccacgtcccg agctacctct ttcctgctgg ctttggagcc
     2521 tgagctcgag gcccggctgg cgcacctggg cgtgcaggta aagggagaag atgaggaaga
     2581 gaacaatttg gaagtacgtg aaaccaaaat taagggtaaa agtgggagat tcttcacagt
     2641 caagctccca gttgctcttg atcctggggc caagatttca gtcattgtgg aaacagtcta
     2701 cacccatgtg cttcatccgt atccaaccca gatcacccag tcagagaaac agtttgtggt
     2761 gtttgagggg aaccattatt tctactctcc ctatccaacg aagacacaaa ccatgcgtgt
     2821 gaagcttgcc tctcgaaatg tggagagcta caccaagctg gggaacccca cgcgctctga
     2881 ggacctactg gattatgggc ctttcagaga tgtgcctgcc tatagtcagg atacttttaa
     2941 agtacattat gagaacaaca gccctttcct gaccatcacc agcatgaccc gagtcattga
     3001 agtctctcac tggggtaata ttgctgtgga agaaaatgtg gacttaaagc acacaggagc
     3061 tgtgcttaag gggcctttct cacgctatga ttaccagaga cagccagata gtggaatatc
     3121 ctccatccgt tcttttaaga ccatccttcc tgctgctgcc caggatgttt attaccggga
     3181 tgagattggc aatgtttcta ccagccacct ccttattttg gatgactctg tagagatgga
     3241 aatccggcct cgcttccctc tctttggcgg gtggaagacc cattacatcg ttggctacaa
     3301 cctcccaagc tatgagtacc tctataattt gggtgaccag tatgcactga agatgaggtt
     3361 tgtggaccat gtgtttgatg aacaagtgat agattctctg actgtgaaga tcatcctgcc
     3421 tgaaggagcc aagaacattg aaattgatag tccctatgaa atcagccgtg ccccagatga
     3481 gctgcactac acctatctgg acacatttgg ccgccctgtg attgttgcct acaagaaaaa
     3541 tctggtagaa cagcacattc aggacattgt ggtccactac acgttcaaca aggtgctcat
     3601 gctgcaggag cccctgctgg tggtggcggc cttctacatc ctgttcttca ccgttatcat
     3661 ctatgttcgg ctggacttct ccatcaccaa ggatccagcc gcagaagcca ggatgaaggt
     3721 agcctgcatc acagagcagg tcttgaccct ggtcaacaag agaataggcc tttaccgtca
     3781 ctttgacgag accgtcaata ggtacaagca atcccgggac atctccaccc tcaacagtgg
     3841 caagaagagc ctggagactg aacacaaggc cttgaccagt gagattgcac tgctgcagtc
     3901 caggctgaag acagagggct ctgatctgtg cgacagagtg agcgaaatgc agaagctgga
     3961 tgcacaggtc aaggagctgg tgctgaagtc ggcggtggag gctgagcgcc tggtggctgg
     4021 caagctcaag aaagacacgt acattgagaa tgagaagctc atctcaggaa agcgccagga
     4081 gctggtcacc aagatcgacc acatcctgga tgccctgtag ggatccgcgg ccgcgaagga
     4141 tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
     4201 agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc ggggtaaact
     4261 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
     4321 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc
     4381 tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc
     4441 atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt
     4501 ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt
     4561 ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct
     4621 acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta
     4681 cgctagacgc caccatggag agcgacgaga gcggcctgcc cgccatggag atcgagtgcc
     4741 gcatcaccgg caccctgaac ggcgtggagt tcgagctggt gggcggcgga gagggcaccc
     4801 ccaagcaggg ccgcatgacc aacaagatga agagcaccaa aggcgccctg accttcagcc
     4861 cctacctgct gagccacgtg atgggctacg gcttctacca cttcggcacc taccccagcg
     4921 gctacgagaa ccccttcctg cacgccatca acaacggcgg ctacaccaac acccgcatcg
     4981 agaagtacga ggacggcggc gtgctgcacg tgagcttcag ctaccgctac gaggccggcc
     5041 gcgtgatcgg cgacttcaag gtggtgggca ccggcttccc cgaggacagc gtgatcttca
     5101 ccgacaagat catccgcagc aacgccaccg tggagcacct gcaccccatg ggcgataacg
     5161 tgctggtggg cagcttcgcc cgcaccttca gcctgcgcga cggcggctac tacagcttcg
     5221 tggtggacag ccacatgcac ttcaagagcg ccatccaccc cagcatcctg cagaacgggg
     5281 gccccatgtt cgccttccgc cgcgtggagg agctgcacag caacaccgag ctgggcatcg
     5341 tggagtacca gcacgccttc aagaccccca tcgccttcgc cagatcccgc gctcagtcgt
     5401 ccaattctgc cgtggacggc accgccggac ccggctccac cggatctcgc taagtcgaca
     5461 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     5521 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     5581 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     5641 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     5701 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     5761 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     5821 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     5881 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     5941 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     6001 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta
     6061 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     6121 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct
     6181 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     6241 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     6301 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     6361 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     6421 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     6481 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     6541 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt
     6601 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     6661 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg
     6721 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6781 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6841 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6901 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6961 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     7021 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     7081 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     7141 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     7201 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7261 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7321 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7381 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7441 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7501 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7561 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     7621 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7681 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7741 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7801 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7861 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7921 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7981 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     8041 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     8101 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     8161 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     8221 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     8281 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8341 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8401 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8461 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8521 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8581 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8641 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8701 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8761 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8821 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8881 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     8941 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     9001 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     9061 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     9121 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     9181 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     9241 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     9301 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca
     9361 agctg
//
