LOCUS       V032983                 8705 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032983
VERSION     V032983
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"
     regulatory      2302..2311
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2311..2352
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     CDS             2353..3624
                     /label="STAMBP(NM_006463)"
                     /note="STAMBP(NM_006463)"
                     /gene="STAMBP"
     promoter        3675..3886
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3899..4167
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4194..4793
                     /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    4800..5388
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5462..5695
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5767..5888
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5907..6042
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6080..6096)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6104..6120
                     /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(6128..6158)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6173..6194
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6482..7070)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7241..8101)
                     /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(8102..8206)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8680..8696
                     /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 cgccaccatg ggtaagccta tccctaaccc tctcctcggt
     2341 ctcgattcta caatgtctga ccatggagat gtgagcctcc cgcccgaaga ccgggtgagg
     2401 gctctctccc agctgggtag tgcggtagag gtgaatgaag acattccacc ccgtcggtac
     2461 ttccgctctg gagttgagat tatccgaatg gcatccattt actctgagga aggcaacatt
     2521 gaacatgcct tcatcctcta taacaagtat atcacgctct ttattgagaa actaccaaaa
     2581 catcgagatt acaaatctgc tgtcattcct gaaaagaaag acacagtaaa gaaattaaag
     2641 gagattgcat ttcccaaagc agaagagctg aaggcagagc tgttaaaacg atataccaaa
     2701 gaatatacag aatataatga agaaaagaag aaggaagcag aggaattggc ccggaacatg
     2761 gccatccagc aagagctgga aaaggaaaaa cagagggtag cacaacagaa gcagcagcaa
     2821 ttggaacagg aacagttcca tgccttcgag gagatgatcc ggaaccagga gctagaaaaa
     2881 gagcgactga aaattgtaca ggagtttggg aaggtagacc ctggcctagg tggcccgcta
     2941 gtgcctgact tggagaagcc ctccttagat gtgttcccca ccttaacagt ctcatccata
     3001 cagccttcag actgtcacac aactgtaagg ccagctaagc cacctgtggt ggacaggtcc
     3061 ttgaaacctg gagcactgag caactcagaa agtattccca caatcgatgg attgcgccat
     3121 gtggtggtgc ctgggcggct gtgcccacag tttctccagt tagccagtgc caacactgcc
     3181 cggggagtgg agacatgtgg aattctctgt ggaaaactga tgaggaatga atttaccatt
     3241 acccatgttc tcatccccaa gcaaagtgct gggtctgatt actgcaacac agagaacgaa
     3301 gaagaacttt tcctcataca ggatcagcag ggcctcatca cactgggctg gattcatact
     3361 caccccacac agaccgcgtt tctctccagt gtcgacctac acactcactg ctcttaccag
     3421 atgatgttgc cagagtcagt agccattgtt tgctccccca agttccagga aactggattc
     3481 tttaaactaa ctgaccatgg actagaggag atttcttcct gtcgccagaa aggatttcat
     3541 ccacacagca aggatccacc tctgttctgt agctgcagcc acgtgactgt tgtggacaga
     3601 gcagtgacca tcacagacct tcgataggga tccgcggccg cgaaggatct gcgatcgctc
     3661 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     3721 ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     3781 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     3841 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg
     3901 ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt
     3961 gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt
     4021 aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag
     4081 actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc
     4141 gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagatgaccg
     4201 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc
     4261 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca
     4321 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     4381 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     4441 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     4501 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     4561 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     4621 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     4681 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     4741 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca
     4801 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     4861 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     4921 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     4981 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     5041 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     5101 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     5161 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     5221 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     5281 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     5341 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta
     5401 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     5461 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct
     5521 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     5581 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     5641 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     5701 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     5761 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5821 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5881 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt
     5941 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     6001 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg
     6061 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6121 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6181 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6241 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6301 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6361 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6421 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6481 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6541 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6601 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6661 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6721 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6781 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6841 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6901 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     6961 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7021 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7081 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7141 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7201 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7261 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7321 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7381 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7441 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7501 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7561 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7621 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7681 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7741 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7801 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7861 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7921 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7981 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8041 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8101 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8161 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8221 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     8281 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     8341 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     8401 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     8461 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     8521 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     8581 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     8641 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca
     8701 agctg
//
