LOCUS       V032376                 8585 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032376
VERSION     V032376
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..3669
                     /label="GABRA1(NM_000806)"
                     /note="GABRA1(NM_000806)"
                     /gene="GABRA1"
     CDS             3670..3699
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHHHHHH"
                     /label="10xHis"
     promoter        3750..3961
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3974..4242
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4275..4673
                     /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    4680..5268
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5342..5575
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5647..5768
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5787..5922
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(5960..5976)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5984..6000
                     /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(6008..6038)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6053..6074
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6362..6950)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7121..7981)
                     /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(7982..8086)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8560..8576
                     /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 catgaggaaa agtccaggtc tgtctgactg tctttgggcc
     2341 tggatcctcc ttctgagcac actgactgga agaagctatg gacagccgtc attacaagat
     2401 gaacttaaag acaataccac tgtcttcacc aggattttgg acagactcct agatggttat
     2461 gacaatcgcc tgagaccagg attgggagag cgtgtaaccg aagtgaagac tgatatcttc
     2521 gtcaccagtt tcggacccgt ttcagaccat gatatggaat atacaataga tgtatttttc
     2581 cgtcaaagct ggaaggatga aaggttaaaa tttaaaggac ctatgacagt cctccggtta
     2641 aataacctaa tggcaagtaa aatctggact ccggacacat ttttccacaa tggaaagaag
     2701 tcagtggccc acaacatgac catgcccaac aaactcctgc ggatcacaga ggatggcacc
     2761 ttgctgtaca ccatgaggct gacagtgaga gctgaatgtc cgatgcattt ggaggacttc
     2821 cctatggatg cccatgcttg cccactaaaa tttggaagtt atgcttatac aagagcagaa
     2881 gttgtttatg aatggaccag agagccagca cgctcagtgg ttgtagcaga agatggatca
     2941 cgtctaaacc agtatgacct tcttggacaa acagtagact ctggaattgt ccagtcaagt
     3001 acaggagaat atgttgttat gaccactcat ttccacttga agagaaagat tggctacttt
     3061 gttattcaaa catacctgcc atgcataatg acagtgattc tctcacaagt ctccttctgg
     3121 ctcaacagag agtctgtacc agcaagaact gtctttggag taacaactgt gctcaccatg
     3181 acaacattga gcatcagtgc cagaaactcc ctccctaagg tggcttatgc aacagctatg
     3241 gattggttta ttgccgtgtg ctatgccttt gtgttctcag ctctgattga gtttgccaca
     3301 gtaaactatt tcactaagag aggttatgca tgggatggca aaagtgtggt tccagaaaag
     3361 ccaaagaaag taaaggatcc tcttattaag aaaaacaaca cttacgctcc aacagcaacc
     3421 agctacaccc ctaatttggc caggggcgac ccgggcttag ccaccattgc taaaagtgca
     3481 accatagaac ctaaagaggt caagcccgaa acaaaaccac cagaacccaa gaaaaccttt
     3541 aacagtgtca gcaaaattga ccgactgtca agaatagcct tcccgctgct atttggaatc
     3601 tttaacttag tctactgggc tacgtattta aacagagagc ctcagctaaa agcccccaca
     3661 ccacatcaac accaccacca tcaccatcat caccaccact gaggatccgc ggccgcgaag
     3721 gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
     3781 gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa
     3841 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
     3901 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
     3961 gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg
     4021 ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc
     4081 gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc
     4141 ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact
     4201 ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc
     4261 tacgctaggc caccatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag
     4321 caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc
     4381 tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt
     4441 gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta
     4501 tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg
     4561 tgcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga
     4621 cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taagtcgaca
     4681 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     4741 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     4801 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     4861 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     4921 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     4981 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     5041 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     5101 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     5161 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     5221 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta
     5281 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     5341 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct
     5401 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     5461 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     5521 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     5581 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     5641 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5701 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5761 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt
     5821 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     5881 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg
     5941 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6001 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6061 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6121 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6181 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6241 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6301 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6361 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6421 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6481 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6541 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6601 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6661 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6721 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6781 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     6841 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6901 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6961 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7021 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7081 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7141 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7201 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7261 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7321 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7381 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7441 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7501 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7561 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7621 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7681 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7741 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7801 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7861 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7921 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7981 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8041 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8101 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     8161 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     8221 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     8281 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     8341 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     8401 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     8461 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     8521 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca
     8581 agctg
//
