LOCUS       V022058                 9235 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022058
VERSION     V022058
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..3277
                     /label="Ech1(NM_016772)"
                     /note="Ech1(NM_016772)"
                     /gene="Ech1"
     CDS             3284..3349
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3394..3605
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3618..3886
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3915..3924
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3945..4607
                     /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"
     CDS             4677..4730
                     /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             4731..5330
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5331..5918
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5992..6225
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6297..6418
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6437..6572
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6610..6626)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6634..6650
                     /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(6658..6688)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6703..6724
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7012..7600)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7771..8631)
                     /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(8632..8736)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9210..9226
                     /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 ctaccgcgat gacagtttcc agtaaactcc gcggcctgct
     2341 gatgcagcag ctgagaggta ccagccagct gtacttcaac atcagccttc gatctctgag
     2401 ctcctctgca caagaggcct ccaagagagc ccccgaggaa gtctcagacc acaactatga
     2461 gtctattcaa gtcacaagtg cccagaagca tgttctccac gtgcagctaa accggccaga
     2521 gaagaggaat gccatgaaca gggctttctg gagggagttg gtggaatgct tccaaaagat
     2581 atccaaagac tcggactgcc gggctgttgt agtctccggt gcaggaaaga tgttcacttc
     2641 aggtattgac ctcatggaca tggcctccga actcatgcag ccctcaggag atgatgcggc
     2701 ccgcatagcc tggtacctcc gtgacctcat cagcaagtac cagaagacct tcactgtcat
     2761 tgagaagtgc cccaagccag tgattgctgc cattcacgga ggctgcattg gcggaggtgt
     2821 ggacctcgtt tctgcctgtg acattcgcta ctgcactcag gatgcttttt tccagatcaa
     2881 ggaggtagat atgggcttgg ctgctgatgt aggaacactg cagagactgc ccaaggtcat
     2941 cgggaaccag agcctggtca acgagctgac cttctctgcc cgcaagatga tggctgacga
     3001 ggccctggac agtgggctgg tcagccgcgt gttccaagat aaggacgcca tgctgaatgc
     3061 agcctttgcc ctggcggctg atatttccag caagagccct gtggctgtgc aggggtcaaa
     3121 aatcaatctc atctactccc gagaccattc tgtggacgag agccttgact acatggccac
     3181 ctggaacatg agcatgctgc agacccagga catcatcaaa tcagtccagg cagccatgga
     3241 gaagagggac acaaaaagca tcaccttctc caagctcgga tccgactaca aagaccatga
     3301 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt gagcggccgc
     3361 gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc
     3421 ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg
     3481 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac
     3541 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa
     3601 cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag
     3661 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa
     3721 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc
     3781 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc
     3841 aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg
     3901 cgcctacgct agacgccacc atggagagcg acgagagcgg cctgcccgcc atggagatcg
     3961 agtgccgcat caccggcacc ctgaacggcg tggagttcga gctggtgggc ggcggagagg
     4021 gcacccccaa gcagggccgc atgaccaaca agatgaagag caccaaaggc gccctgacct
     4081 tcagccccta cctgctgagc cacgtgatgg gctacggctt ctaccacttc ggcacctacc
     4141 ccagcggcta cgagaacccc ttcctgcacg ccatcaacaa cggcggctac accaacaccc
     4201 gcatcgagaa gtacgaggac ggcggcgtgc tgcacgtgag cttcagctac cgctacgagg
     4261 ccggccgcgt gatcggcgac ttcaaggtgg tgggcaccgg cttccccgag gacagcgtga
     4321 tcttcaccga caagatcatc cgcagcaacg ccaccgtgga gcacctgcac cccatgggcg
     4381 ataacgtgct ggtgggcagc ttcgcccgca ccttcagcct gcgcgacggc ggctactaca
     4441 gcttcgtggt ggacagccac atgcacttca agagcgccat ccaccccagc atcctgcaga
     4501 acgggggccc catgttcgcc ttccgccgcg tggaggagct gcacagcaac accgagctgg
     4561 gcatcgtgga gtaccagcac gccttcaaga cccccatcgc cttcgccaga tcccgcgctc
     4621 agtcgtccaa ttctgccgtg gacggcaccg ccggacccgg ctccaccgga tctcgcgagg
     4681 gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct atgaccgagt
     4741 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     4801 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     4861 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     4921 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     4981 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     5041 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     5101 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     5161 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     5221 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     5281 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga aatcaacctc
     5341 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     5401 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     5461 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     5521 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     5581 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     5641 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     5701 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg
     5761 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     5821 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctc cgccttcgcc
     5881 ctcagacgag tcggatctcc ctttggccgc ctccccgcct ggtaccttta agaccaatga
     5941 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     6001 taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct ctctggttag
     6061 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     6121 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     6181 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     6241 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact
     6301 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6361 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6421 atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     6481 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     6541 ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg gcccaaattc
     6601 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6661 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6721 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6781 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6841 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6901 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6961 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     7021 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7081 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7141 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7201 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7261 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7321 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7381 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7441 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7501 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7561 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7621 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7681 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7741 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7801 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7861 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7921 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7981 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8041 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8101 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8161 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8221 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8281 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8341 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8401 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8461 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8521 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8581 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8641 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8701 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8761 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     8821 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     8881 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     8941 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     9001 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     9061 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     9121 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     9181 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctg
//
