LOCUS       V033973                 9515 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033973
VERSION     V033973
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..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..4347
                     /label="GFPT2(NM_005110)"
                     /note="GFPT2(NM_005110)"
                     /gene="GFPT2"
     CDS             4354..4380
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4384..4410
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4414..4440
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        4485..4696
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4709..4977
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5004..5603
                     /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    5610..6198
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6272..6505
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6577..6698
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6717..6852
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6890..6906)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6914..6930
                     /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(6938..6968)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6983..7004
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7292..7880)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8051..8911)
                     /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(8912..9016)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9490..9506
                     /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 attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catgtgcgga atctttgcct acatgaacta cagagtcccc
     2341 cggacgagga aggagatctt cgaaaccctc atcaagggcc tgcagcggct ggagtacaga
     2401 ggctacgact cggcaggtgt ggcgatcgat gggaataatc acgaagtcaa agaaagacac
     2461 attcagctgg tcaagaaaag ggggaaagtc aaggctctcg atgaagaact ttacaaacaa
     2521 gacagcatgg acttaaaagt ggagtttgag acacacttcg gcattgccca cacgcgctgg
     2581 gccacccacg gggtccccag tgctgtcaac agccaccctc agcgctcaga caaaggcaac
     2641 gaatttgttg tcatccacaa tgggatcatc acaaattaca aagatctgag gaaatttctg
     2701 gaaagcaaag gctacgagtt tgagtcagaa acagatacag agaccatcgc caagctgatt
     2761 aaatatgtgt tcgacaacag agaaactgag gacattacgt tttcaacgtt ggtcgagaga
     2821 gtcattcagc agttggaagg tgcattcgcg ctggttttca agagtgtcca ctacccagga
     2881 gaagccgttg ccacacggag aggcagcccc ctgctcatcg gagtccggag caaatacaag
     2941 ctctccacag aacagatccc tatcttatac aggacgtgca ctctggagaa tgtgaagaat
     3001 atctgtaaga cacggatgaa gaggctggac agctccgcct gcctgcatgc tgtgggcgac
     3061 aaggccgtgg aattcttctt tgcttctgat gcaagcgcta tcatagagca caccaaccgg
     3121 gtcatcttcc tggaggacga tgacatcgcc gcagtggctg atgggaaact ctccattcac
     3181 cgggtcaagc gctcggccag tgatgaccca tctcgagcca tccagacctt gcagatggaa
     3241 ctgcagcaaa tcatgaaagg taacttcagt gcgtttatgc agaaggagat cttcgaacag
     3301 ccagaatcag ttttcaatac tatgagaggt cgggtgaatt ttgaaaccaa cacagtgctc
     3361 ctgggtggct tgaaggacca cttgaaggag attcgacgat gccgacggct catcgtgatt
     3421 ggctgtggaa ccagctacca cgctgccgtg gctacgcggc aagttttgga ggaactgact
     3481 gagcttcctg tgatggttga acttgctagt gattttctgg acaggaacac acctgtgttc
     3541 agggatgacg tttgcttttt catcagccag tcaggcgaga ccgcggacac cctcctggcg
     3601 ctgcgctact gtaaggaccg cggcgctctc accgtgggcg tcaccaacac cgtgggcagc
     3661 tccatctctc gcgagaccga ctgcggcgtc cacatcaacg cagggccgga gatcggcgtg
     3721 gccagcacca aggcttatac cagtcagttc atctctctgg tgatgtttgg tttgatgatg
     3781 tctgaagacc gaatttcact acaaaacagg aggcaagaga tcatccgtgg cttgagatct
     3841 ttacctgagc tgatcaagga agtgctgtct ctggaggaga agatccacga cttggccctg
     3901 gagctctaca cgcagagatc gctgctggtg atggggcggg gctacaacta tgccacctgc
     3961 ctggaaggag ccctgaaaat taaagagata acctacatgc actcagaagg catcctggct
     4021 ggggagctga agcacgggcc cctggcactg attgacaagc agatgcccgt catcatggtc
     4081 attatgaagg atccttgctt cgccaaatgc cagaacgccc tgcagcaagt cacggcccgc
     4141 cagggtcgcc ccattatact gtgctccaag gacgatactg aaagttccaa gtttgcgtat
     4201 aagacaattg agctgcccca cactgtggac tgcctccagg gcatcctgag cgtgattccg
     4261 ctgcagctgc tgtccttcca cctggctgtt ctccgaggat atgacgttga cttccccaga
     4321 aatctggcca agtctgtaac tgtggaagga tcctacccat acgacgtacc tgattatgca
     4381 acttatcctt atgatgtacc tgattatgct agctatcctt atgatgtccc agactacgca
     4441 taggcggccg cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc
     4501 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag
     4561 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     4621 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     4681 tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg
     4741 ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg
     4801 tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct
     4861 ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac
     4921 cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa
     4981 gctgtgaccg gcgcctacgc tagatgaccg agtacaagcc cacggtgcgc ctcgccaccc
     5041 gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
     5101 cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct
     5161 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
     5221 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
     5281 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
     5341 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg
     5401 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
     5461 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
     5521 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
     5581 tgacccgcaa gcccggtgcc tgagtcgaca atcaacctct ggattacaaa atttgtgaaa
     5641 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     5701 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     5761 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     5821 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     5881 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     5941 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     6001 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     6061 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     6121 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     6181 tttgggccgc ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat
     6241 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaaa
     6301 taagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     6361 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     6421 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     6481 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     6541 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     6601 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6661 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     6721 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6781 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     6841 tttttggagg cctagacttt tgcagagacg gcccaaattc gtaatcatgg tcatagctgt
     6901 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6961 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     7021 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     7081 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     7141 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     7201 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     7261 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     7321 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     7381 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     7441 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     7501 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7561 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7621 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7681 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     7741 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7801 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7861 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7921 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7981 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     8041 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     8101 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     8161 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     8221 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     8281 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     8341 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     8401 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     8461 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     8521 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8581 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8641 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8701 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8761 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8821 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8881 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8941 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     9001 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     9061 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     9121 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     9181 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     9241 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     9301 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
     9361 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     9421 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     9481 cacgacgttg taaaacgacg gccagtgcca agctg
//
