LOCUS       V032175                 9065 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032175
VERSION     V032175
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..3897
                     /label="SLC1A4(NM_003038)"
                     /note="SLC1A4(NM_003038)"
                     /gene="SLC1A4"
     CDS             3904..3930
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3934..3960
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3964..3990
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        4035..4246
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4259..4527
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4554..5153
                     /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    5160..5748
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5822..6055
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6127..6248
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6267..6402
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6440..6456)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6464..6480
                     /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(6488..6518)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6533..6554
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6842..7430)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7601..8461)
                     /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(8462..8566)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9040..9056
                     /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 catggagaag agcaacgaga ccaacggcta ccttgacagc
     2341 gctcaggcgg ggcctgcggc cgggcccgga gctccgggga ccgcggcggg acgcgcacgg
     2401 cgttgcgcgg gcttcctgcg gcgccaagcg ctggtgctgc tcaccgtgtc cggggtgctg
     2461 gcgggcgcgg gcctgggcgc ggcgttgcgc gggctcagcc tgagccgcac gcaggtcacc
     2521 tacctggcct tccccggcga gatgctgctc cgcatgctgc gcatgatcat cctgccgctg
     2581 gtggtctgca gcctggtgtc gggcgccgcc tcgctcgatg ccagctgcct cgggcgtctg
     2641 ggcggcatcg ctgtcgccta ctttggcctc accacactga gtgcctcggc gctcgccgtg
     2701 gccttggcgt tcatcatcaa gccaggatcc ggtgcgcaga cccttcagtc cagcgacctg
     2761 gggctggagg actcggggcc tcctcctgtc cccaaagaga cggtggactc tttcctcgac
     2821 ctggccagaa acctgtttcc ctccaatctt gtggttgcag ctttccgtac gtatgcaacc
     2881 gattataaag tcgtgaccca gaacagcagc tctggaaatg taacccatga aaagatcccc
     2941 ataggcactg agatagaagg gatgaacatt ttaggattgg tcctgtttgc tctggtgtta
     3001 ggagtggcct taaagaaact aggctccgaa ggagaagacc tcatccgttt cttcaattcc
     3061 ctcaacgagg cgacgatggt gctggtgtcc tggattatgt ggtacgtacc tgtgggcatc
     3121 atgttccttg ttggaagcaa gatcgtggaa atgaaagaca tcatcgtgct ggtgaccagc
     3181 ctggggaaat acatcttcgc atctatattg ggccatgtta ttcatggagg aattgttctg
     3241 ccacttattt attttgtttt cacacgaaaa aacccattca gattcctcct gggcctcctc
     3301 gccccatttg cgacagcatt tgctacctgc tccagctcag cgacccttcc ctctatgatg
     3361 aagtgcattg aagagaacaa tggtgtggac aagaggatca gcaggtttat tctccccatc
     3421 ggggccaccg tgaacatgga cggagcagcc atcttccagt gtgtggccgc ggtgttcatt
     3481 gcgcaactca acaacgtaga gctcaacgca ggacagattt tcaccattct agtgactgcc
     3541 acagcgtcca gtgttggagc agcaggcgtg ccagctggag gggtcctcac cattgccatt
     3601 atcctggagg ccattgggct gcctactcat gacctgcctc tgatcctggc tgtggactgg
     3661 attgtggacc ggaccaccac ggtggtgaat gtggaagggg atgccctggg tgcaggcatt
     3721 ctccaccacc tgaatcagaa ggcaacaaag aaaggcgagc aggaacttgc tgaggtgaaa
     3781 gtggaagcca tccccaactg caagtctgag gaggagacat cgcccctggt gacacaccag
     3841 aaccccgctg gccccgtggc cagtgcccca gaactggaat ccaaggagtc ggttctggga
     3901 tcctacccat acgacgtacc tgattatgca acttatcctt atgatgtacc tgattatgct
     3961 agctatcctt atgatgtccc agactacgca taggcggccg cgaaggatct gcgatcgctc
     4021 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     4081 ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     4141 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     4201 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg
     4261 ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt
     4321 gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt
     4381 aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag
     4441 actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc
     4501 gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagatgaccg
     4561 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc
     4621 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca
     4681 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     4741 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     4801 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     4861 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     4921 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     4981 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     5041 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     5101 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca
     5161 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     5221 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     5281 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     5341 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     5401 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     5461 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     5521 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     5581 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     5641 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     5701 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta
     5761 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     5821 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct
     5881 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     5941 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     6001 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     6061 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     6121 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     6181 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     6241 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt
     6301 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     6361 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg
     6421 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6481 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6541 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6601 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6661 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6721 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6781 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6841 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6901 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6961 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7021 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7081 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7141 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7201 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7261 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     7321 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7381 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7441 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7501 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7561 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7621 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7681 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7741 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7801 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7861 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7921 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7981 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8041 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8101 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8161 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8221 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8281 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8341 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8401 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8461 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8521 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8581 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     8641 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     8701 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     8761 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     8821 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     8881 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     8941 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     9001 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca
     9061 agctg
//
