LOCUS       V032795                 9506 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032795
VERSION     V032795
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..4338
                     /label="SLC20A1(NM_005415)"
                     /note="SLC20A1(NM_005415)"
                     /gene="SLC20A1"
     CDS             4345..4371
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4375..4401
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4405..4431
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        4476..4687
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4700..4968
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4995..5594
                     /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    5601..6189
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6263..6496
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6568..6689
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6708..6843
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6881..6897)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6905..6921
                     /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(6929..6959)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6974..6995
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7283..7871)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8042..8902)
                     /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(8903..9007)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9481..9497
                     /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 catggcaacg ctgattacca gtactacagc tgctaccgcc
     2341 gcttctggtc ctttggtgga ctacctatgg atgctcatcc tgggcttcat tattgcattt
     2401 gtcttggcat tctccgtggg agccaatgat gtagcaaatt cttttggtac agctgtgggc
     2461 tcaggtgtag tgaccctgaa gcaagcctgc atcctagcta gcatctttga aacagtgggc
     2521 tctgtcttac tgggggccaa agtgagcgaa accatccgga agggcttgat tgacgtggag
     2581 atgtacaact cgactcaagg gctgctgatg gccggctcag tcagtgctat gtttggttct
     2641 gctgtgtggc aactcgtggc ttcgtttttg aagctcccta tttctggaac ccattgtatt
     2701 gttggtgcaa ctattggttt ctccctcgtg gcaaaggggc aggagggtgt caagtggtct
     2761 gaactgataa aaattgtgat gtcttggttc gtgtccccac tgctttctgg aattatgtct
     2821 ggaattttat tcttcctggt tcgtgcattc atcctccata aggcagatcc agttcctaat
     2881 ggtttgcgag ctttgccagt tttctatgcc tgcacagttg gaataaacct cttttccatc
     2941 atgtatactg gagcaccgtt gctgggcttt gacaaacttc ctctgtgggg taccatcctc
     3001 atctcggtgg gatgtgcagt tttctgtgcc cttatcgtct ggttctttgt atgtcccagg
     3061 atgaagagaa aaattgaacg agaaataaag tgtagtcctt ctgaaagccc cttaatggaa
     3121 aaaaagaata gcttgaaaga agaccatgaa gaaacaaagt tgtctgttgg tgatattgaa
     3181 aacaagcatc ctgtttctga ggtagggcct gccactgtgc ccctccaggc tgtggtggag
     3241 gagagaacag tctcattcaa acttggagat ttggaggaag ctccagagag agagaggctt
     3301 cccagcgtgg acttgaaaga ggaaaccagc atagatagca ccgtgaatgg tgcagtgcag
     3361 ttgcctaatg ggaaccttgt ccagttcagt caagccgtca gcaaccaaat aaactccagt
     3421 ggccactacc agtatcacac cgtgcataag gattccggcc tgtacaaaga gctactccat
     3481 aaattacatc ttgccaaggt gggagattgc atgggagact ccggtgacaa acccttaagg
     3541 cgcaataata gctatacttc ctataccatg gcaatatgtg gcatgcctct ggattcattc
     3601 cgtgccaaag aaggtgaaca gaagggcgaa gaaatggaga agctgacatg gcctaatgca
     3661 gactccaaga agcgaattcg aatggacagt tacaccagtt actgcaatgc tgtgtctgac
     3721 cttcactcag catctgagat agacatgagt gtcaaggcag agatgggtct aggtgacaga
     3781 aaaggaagta atggctctct agaagaatgg tatgaccagg ataagcctga agtctctctc
     3841 ctcttccagt tcctgcagat ccttacagcc tgctttgggt cattcgccca tggtggcaat
     3901 gacgtaagca atgccattgg gcctctggtt gctttatatt tggtttatga cacaggagat
     3961 gtttcttcaa aagtggcaac accaatatgg cttctactct atggtggtgt tggtatctgt
     4021 gttggtctgt gggtttgggg aagaagagtt atccagacca tggggaagga tctgacaccg
     4081 atcacaccct ctagtggctt cagtattgaa ctggcatctg ccctcactgt ggtgattgca
     4141 tcaaatattg gccttcccat cagtacaaca cattgtaaag tgggctctgt tgtgtctgtt
     4201 ggctggctcc ggtccaagaa ggctgttgac tggcgtctct ttcgtaacat ttttatggcc
     4261 tggtttgtca cagtccccat ttctggagtt atcagtgctg ccatcatggc aatcttcaga
     4321 tatgtcatcc tcagaatggg atcctaccca tacgacgtac ctgattatgc aacttatcct
     4381 tatgatgtac ctgattatgc tagctatcct tatgatgtcc cagactacgc ataggcggcc
     4441 gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     4501 ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg
     4561 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     4621 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     4681 aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg
     4741 aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg
     4801 aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc
     4861 gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc
     4921 tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc
     4981 ggcgcctacg ctagatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     5041 tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     5101 ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     5161 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     5221 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     5281 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     5341 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     5401 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     5461 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     5521 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     5581 agcccggtgc ctgagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     5641 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     5701 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     5761 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     5821 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     5881 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     5941 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     6001 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     6061 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     6121 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     6181 cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     6241 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg
     6301 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     6361 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     6421 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     6481 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     6541 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     6601 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6661 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     6721 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     6781 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     6841 gcctagactt ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt
     6901 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6961 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     7021 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     7081 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     7141 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     7201 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     7261 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     7321 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     7381 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     7441 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     7501 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7561 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7621 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7681 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     7741 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7801 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7861 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7921 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7981 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     8041 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     8101 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     8161 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     8221 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     8281 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     8341 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     8401 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     8461 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     8521 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     8581 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8641 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8701 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8761 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8821 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8881 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8941 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     9001 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     9061 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     9121 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     9181 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     9241 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     9301 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
     9361 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     9421 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     9481 gtaaaacgac ggccagtgcc aagctg
//
