LOCUS       V036998                 9495 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036998
VERSION     V036998
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             2317..3711
                     /label="TACR3(NM_001059)"
                     /note="TACR3(NM_001059)"
                     /gene="TACR3"
     CDS             3718..3783
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3848..4059
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4072..4340
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4369..4378
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4399..5061
                     /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             5131..5184
                     /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             5185..5583
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     misc_feature    5590..6178
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6252..6485
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6557..6678
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6697..6832
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6870..6886)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6894..6910
                     /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(6918..6948)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6963..6984
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7272..7860)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8031..8891)
                     /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(8892..8996)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9470..9486
                     /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 ctgatacgaa ctcggaattc gccaccatgg ccactctccc agcagcagaa
     2341 acctggatag acgggggtgg aggcgtgggt gcagacgccg tgaacctgac cgcctcgcta
     2401 gctgccgggg cggccacggg ggcagttgag actgggtggc tgcaactgct ggaccaagct
     2461 ggcaacctct cctcctcccc ttccgcgctg ggactgcctg tggcttcccc cgcgccctcc
     2521 cagccctggg ccaacctcac caaccagttc gtgcagccgt cctggcgcat cgcgctctgg
     2581 tccctggcgt atggtgtggt ggtggcagtg gcagttttgg gaaatctcat cgtcatctgg
     2641 atcatcctgg cccacaagcg catgaggact gtcaccaact acttccttgt gaacctggct
     2701 ttctccgacg cctccatggc cgccttcaac acgttggtca atttcatcta cgcgcttcat
     2761 agcgagtggt actttggcgc caactactgc cgcttccaga acttctttcc tatcacagct
     2821 gtgttcgcca gcatctactc catgacggcc attgcggtgg acaggtatat ggctattatt
     2881 gatcccttga aacccagact gtctgctaca gcaaccaaga ttgtcattgg aagtatttgg
     2941 attctagcat ttctacttgc cttccctcag tgtctttatt ccaaaaccaa agtcatgcca
     3001 ggccgtactc tctgctttgt gcaatggcca gaaggtccca aacaacattt cacttaccat
     3061 attatcgtca ttatactggt gtactgtttc ccattgctca tcatgggtat tacatacacc
     3121 attgttggaa ttactctctg gggaggagaa atcccaggag atacctgtga caagtatcat
     3181 gagcagctaa aggccaaaag aaaggttgtc aaaatgatga ttattgttgt catgacattt
     3241 gctatctgct ggctgcccta tcatatttac ttcattctca ctgcaatcta tcaacaacta
     3301 aatagatgga aatacatcca gcaggtctac ctggctagct tttggctggc aatgagctca
     3361 accatgtaca atcccatcat ctactgctgt ctgaataaaa gatttcgagc tggcttcaag
     3421 agagcatttc gctggtgtcc tttcatcaaa gtttccagct atgatgagct agagctcaag
     3481 accaccaggt ttcatccaaa ccggcaaagc agtatgtaca ccgtgaccag aatggagtcc
     3541 atgacagtcg tgtttgaccc caacgatgca gacaccacca ggtccagtcg gaagaaaaga
     3601 gcaacgccaa gagacccaag tttcaatggc tgctctcgca ggaattccaa atctgcctcc
     3661 gccacttcaa gtttcataag ctcaccctat acctctgtgg atgaatattc tctcgaggac
     3721 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac
     3781 aagtagggat ccctgcgata tccagcgcgg ccgcgaagga tctgcgatcg ctccggtgcc
     3841 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
     3901 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     3961 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     4021 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc
     4081 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc
     4141 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta
     4201 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc
     4261 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct
     4321 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagacgc caccatggag
     4381 agcgacgaga gcggcctgcc cgccatggag atcgagtgcc gcatcaccgg caccctgaac
     4441 ggcgtggagt tcgagctggt gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc
     4501 aacaagatga agagcaccaa aggcgccctg accttcagcc cctacctgct gagccacgtg
     4561 atgggctacg gcttctacca cttcggcacc taccccagcg gctacgagaa ccccttcctg
     4621 cacgccatca acaacggcgg ctacaccaac acccgcatcg agaagtacga ggacggcggc
     4681 gtgctgcacg tgagcttcag ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag
     4741 gtggtgggca ccggcttccc cgaggacagc gtgatcttca ccgacaagat catccgcagc
     4801 aacgccaccg tggagcacct gcaccccatg ggcgataacg tgctggtggg cagcttcgcc
     4861 cgcaccttca gcctgcgcga cggcggctac tacagcttcg tggtggacag ccacatgcac
     4921 ttcaagagcg ccatccaccc cagcatcctg cagaacgggg gccccatgtt cgccttccgc
     4981 cgcgtggagg agctgcacag caacaccgag ctgggcatcg tggagtacca gcacgccttc
     5041 aagaccccca tcgccttcgc cagatcccgc gctcagtcgt ccaattctgc cgtggacggc
     5101 accgccggac ccggctccac cggatctcgc gagggcagag gaagtcttct aacatgcggt
     5161 gacgtggagg agaatcccgg ccctatggcc aagcctttgt ctcaagaaga atccaccctc
     5221 attgaaagag caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc
     5281 agcgcagctc tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact
     5341 gggggacctt gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac
     5401 ctgacttgta tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg
     5461 tgccgacagg tgcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat
     5521 ggacagccga cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc
     5581 taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     5641 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     5701 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     5761 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     5821 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     5881 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     5941 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     6001 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     6061 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     6121 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     6181 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     6241 aaagggggga ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg
     6301 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     6361 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     6421 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     6481 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     6541 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     6601 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6661 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt
     6721 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     6781 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt
     6841 tgcagagacg gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     6901 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     6961 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     7021 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     7081 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     7141 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     7201 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     7261 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     7321 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     7381 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7441 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     7501 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7561 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7621 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7681 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     7741 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7801 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7861 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7921 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7981 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     8041 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     8101 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     8161 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     8221 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     8281 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     8341 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     8401 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8461 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8521 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8581 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8641 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8701 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8761 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8821 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8881 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8941 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     9001 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     9061 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     9121 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     9181 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     9241 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     9301 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
     9361 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     9421 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     9481 gccagtgcca agctg
//
