LOCUS       V036495                 9239 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036495
VERSION     V036495
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             2290..3732
                     /label="Akt2(NM_001110208)"
                     /note="Akt2(NM_001110208)"
                     /gene="Akt2"
     promoter        3782..3993
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4006..4274
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4302..5327
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /note="confers resistance to hygromycin"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /transl_table=1
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E*"
                     /label="HygR"
     misc_feature    5334..5922
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5996..6229
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6301..6422
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6441..6576
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6614..6630)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6638..6654
                     /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(6662..6692)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6707..6728
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7016..7604)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7775..8635)
                     /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(8636..8740)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9214..9230
                     /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 agagccacca tgaatgaggt atctgtcatc aaagaaggct ggctccacaa acgtggtgaa
     2341 tacatcaaga cctggaggcc acggtacttc cttctgaaga gtgatggatc tttcattggg
     2401 tataaggaga ggcccgaggc ccctgaccag accttacccc ccctgaacaa tttctctgta
     2461 gcagaatgcc agctgatgaa gactgagagg ccacgaccca acacctttgt catacgctgc
     2521 ctgcagtgga ccacagtcat cgagaggacc ttccatgtag actctccaga tgagagggaa
     2581 gagtggatgc gggctatcca gatggtcgcc aacagtctga agcagcgggg cccaggtgag
     2641 gacgccatgg attacaagtg tggctccccc agtgactctt ccacatctga gatgatggag
     2701 gtagctgtca acaaggcacg ggccaaagtg accatgaatg acttcgatta tctcaaactc
     2761 ctcggcaagg gcaccttcgg caaggtcatt ctggttcgag agaaggccac tggccgctat
     2821 tatgccatga agatcctgcg caaggaggtc atcattgcaa aggatgaagt cgcccacaca
     2881 gtcacagaga gccgggttct gcagaatacc aggcacccct tccttacagc cctcaagtat
     2941 gccttccaga cccatgaccg cctatgcttt gtgatggagt atgccaacgg gggtgagctg
     3001 tttttccacc tctctcggga gcgagtcttc acggaggatc gggcgcgctt ttatggagca
     3061 gagattgtgt cagctctgga gtatttgcac tcgagagatg tggtgtaccg tgacatcaag
     3121 ctggaaaacc ttatgttgga caaagatggc cacatcaaga tcactgactt tggcttgtgc
     3181 aaagagggca tcagtgatgg agccaccatg aaaaccttct gtggtacccc ggagtacttg
     3241 gcgcctgagg tgctagagga caatgactat gggcgagcag tggactggtg ggggctgggt
     3301 gtggtcatgt atgagatgat gtgtggccgc ctgccattct acaaccagga ccacgagcgc
     3361 ctctttgagc tcattcttat ggaggagatc cgcttcccgc gcacactcgg gccagaggcc
     3421 aagtccctgc tggctggact gctgaagaag gacccaaagc agaggctcgg cggaggtccc
     3481 agtgatgcga aggaggtcat ggagcataga ttcttcctca gcatcaactg gcaggacgtg
     3541 gtacagaaaa agctcctgcc acccttcaaa cctcaggtca cttcagaagt ggacacaagg
     3601 tactttgatg acgagttcac cgcccagtcc atcacaatca cacccccaga ccgatatgac
     3661 agcctggacc cgctggaact ggaccagcgg acgcacttcc cccagttctc ctactcagcc
     3721 agcatccgag agtgaggatc cgcggccgca aggatctgcg atcgctccgg tgcccgtcag
     3781 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     3841 acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     3901 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     3961 ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct
     4021 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
     4081 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
     4141 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
     4201 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg
     4261 cgccgttaca gatccaagct gtgaccggcg cctacgtcga catgaaaaag cctgaactca
     4321 ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc
     4381 agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg
     4441 tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact
     4501 ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc agcgagagcc
     4561 tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg
     4621 aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc
     4681 ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat
     4741 ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg
     4801 acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg
     4861 actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg
     4921 acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat
     4981 acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc
     5041 gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc
     5101 tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag
     5161 cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta
     5221 cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg
     5281 atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaataggtc gacaatcaac
     5341 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     5401 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     5461 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     5521 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     5581 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     5641 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     5701 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     5761 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     5821 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     5881 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggtacc tttaagacca
     5941 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     6001 gggctaattc actcccaacg aaaataagat ctgctttttg cttgtactgg gtctctctgg
     6061 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     6121 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     6181 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
     6241 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     6301 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     6361 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6421 tatcatgtct ggctctagct atcccgcccc taactccgcc cagttccgcc cattctccgc
     6481 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     6541 tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga gacggcccaa
     6601 attcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     6661 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6721 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6781 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6841 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6901 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6961 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     7021 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     7081 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     7141 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     7201 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     7261 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     7321 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7381 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7441 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7501 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     7561 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     7621 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     7681 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     7741 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     7801 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     7861 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     7921 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7981 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     8041 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     8101 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     8161 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     8221 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     8281 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     8341 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     8401 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     8461 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     8521 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     8581 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     8641 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     8701 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     8761 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     8821 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     8881 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     8941 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     9001 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     9061 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     9121 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     9181 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagctg
//
