LOCUS       V026314                 9335 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026314
VERSION     V026314
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    1804..1921
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2011..2214
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2303..3172
                     /label="CD274(NM_014143)"
                     /note="CD274(NM_014143)"
                     /gene="CD274"
     CDS             3179..3244
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3288..3499
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3512..3780
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3809..3818
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3839..4501
                     /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             4571..4624
                     /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             4631..5425
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     misc_feature    5432..6020
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6092..6325
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6397..6518
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6537..6672
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6710..6726)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6734..6750
                     /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(6758..6788)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6803..6824
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7112..7700)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7871..8731)
                     /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(8732..8836)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9310..9326
                     /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 ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc gaattcgcca ccatgaggat atttgctgtc tttatattca tgacctactg
     2341 gcatttgctg aacgcattta ctgtcacggt tcccaaggac ctatatgtgg tagagtatgg
     2401 tagcaatatg acaattgaat gcaaattccc agtagaaaaa caattagacc tggctgcact
     2461 aattgtctat tgggaaatgg aggataagaa cattattcaa tttgtgcatg gagaggaaga
     2521 cctgaaggtt cagcatagta gctacagaca gagggcccgg ctgttgaagg accagctctc
     2581 cctgggaaat gctgcacttc agatcacaga tgtgaaattg caggatgcag gggtgtaccg
     2641 ctgcatgatc agctatggtg gtgccgacta caagcgaatt actgtgaaag tcaatgcccc
     2701 atacaacaaa atcaaccaaa gaattttggt tgtggatcca gtcacctctg aacatgaact
     2761 gacatgtcag gctgagggct accccaaggc cgaagtcatc tggacaagca gtgaccatca
     2821 agtcctgagt ggtaagacca ccaccaccaa ttccaagaga gaggagaagc ttttcaatgt
     2881 gaccagcaca ctgagaatca acacaacaac taatgagatt ttctactgca cttttaggag
     2941 attagatcct gaggaaaacc atacagctga attggtcatc ccagaactac ctctggcaca
     3001 tcctccaaat gaaaggactc acttggtaat tctgggagcc atcttattat gccttggtgt
     3061 agcactgaca ttcatcttcc gtttaagaaa agggagaatg atggatgtga aaaaatgtgg
     3121 catccaagat acaaactcaa agaagcaaag tgatacacat ttggaggaga cgggatccga
     3181 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga
     3241 caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac
     3301 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag
     3361 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     3421 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     3481 gtttgccgcc agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg
     3541 ccgccctacc tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg
     3601 tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg
     3661 cctttgtccg gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct
     3721 gaccctgctt gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc
     3781 caagctgtga ccggcgccta cgctagacgc caccatggag agcgacgaga gcggcctgcc
     3841 cgccatggag atcgagtgcc gcatcaccgg caccctgaac ggcgtggagt tcgagctggt
     3901 gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc aacaagatga agagcaccaa
     3961 aggcgccctg accttcagcc cctacctgct gagccacgtg atgggctacg gcttctacca
     4021 cttcggcacc taccccagcg gctacgagaa ccccttcctg cacgccatca acaacggcgg
     4081 ctacaccaac acccgcatcg agaagtacga ggacggcggc gtgctgcacg tgagcttcag
     4141 ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag gtggtgggca ccggcttccc
     4201 cgaggacagc gtgatcttca ccgacaagat catccgcagc aacgccaccg tggagcacct
     4261 gcaccccatg ggcgataacg tgctggtggg cagcttcgcc cgcaccttca gcctgcgcga
     4321 cggcggctac tacagcttcg tggtggacag ccacatgcac ttcaagagcg ccatccaccc
     4381 cagcatcctg cagaacgggg gccccatgtt cgccttccgc cgcgtggagg agctgcacag
     4441 caacaccgag ctgggcatcg tggagtacca gcacgccttc aagaccccca tcgccttcgc
     4501 cagatcccgc gctcagtcgt ccaattctgc cgtggacggc accgccggac ccggctccac
     4561 cggatctcgc gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg
     4621 ccctgccacc atggctgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4681 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4741 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4801 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4861 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4921 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4981 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5041 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5101 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     5161 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5221 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5281 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5341 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5401 tcgccttctt gacgagttct tctgagtcga caatcaacct ctggattaca aaatttgtga
     5461 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     5521 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     5581 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     5641 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     5701 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     5761 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     5821 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     5881 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     5941 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     6001 cctttgggcc gcctccccgc ggtaccttta agaccaatga cttacaaggc agctgtagat
     6061 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     6121 taagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     6181 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     6241 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     6301 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     6361 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     6421 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6481 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     6541 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6601 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     6661 tttttggagg cctagacttt tgcagagacg gcccaaattc gtaatcatgg tcatagctgt
     6721 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6781 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6841 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6901 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6961 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     7021 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     7081 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     7141 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     7201 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     7261 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     7321 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7381 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7441 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7501 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     7561 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7621 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7681 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7741 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7801 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7861 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7921 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7981 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     8041 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     8101 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     8161 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     8221 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     8281 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     8341 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8401 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8461 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8521 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8581 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8641 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8701 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8761 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8821 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     8881 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     8941 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     9001 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     9061 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     9121 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
     9181 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     9241 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     9301 cacgacgttg taaaacgacg gccagtgcca agctg
//
