LOCUS       V041896                10176 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041896
VERSION     V041896
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /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    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     promoter        2221..2432
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     intron          2433..3371
                     /note="intron upstream of the start codon of human EF-1α"
                     /label="EF-1α intron A"
     regulatory      3387..3396
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3396..3425
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             3426..4853
                     /label="Sec61a1(NM_016906)"
                     /note="Sec61a1(NM_016906)"
                     /gene="Sec61a1"
     misc_feature    4970..5519
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES"
     CDS             5522..6121
                     /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    6145..6733
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6940..7573
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7701..7717)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7725..7741
                     /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(7749..7779)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7794..7815
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8103..8691)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8862..9722)
                     /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(9723..9827)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9875..10009
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgatgaggcc ctttcgtctt cactcgaggt gcccgtcagt
     2221 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     2281 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     2341 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     2401 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
     2461 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt
     2521 acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg
     2581 cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc
     2641 gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca
     2701 tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg
     2761 cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc
     2821 gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg
     2881 gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta
     2941 tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat
     3001 ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc
     3061 gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat
     3121 gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga
     3181 gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt
     3241 gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc
     3301 tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc
     3361 ttccatttca ggtgtcgtga acgcgtgcca ccatggaaca aaaactcatc tcagaagagg
     3421 atctgatggc gatcaaattt ctggaagtca tcaagccatt ctgtgtcatc ttgccggaaa
     3481 ttcagaagcc agagagaaag attcagttta aggagaaagt tctgtggaca gccatcaccc
     3541 tcttcatctt cttagtatgc tgtcagatcc ccctgtttgg catcatgtct tcagactcag
     3601 ctgacccgtt ctactggatg cgagtgattc tggcctctaa cagaggtacc ttaatggagc
     3661 tgggcatctc tcctattgtc acctccggtc ttatcatgca gctcttagct ggtgccaaga
     3721 taattgaagt tggggacacc ccgaaagacc gagctctctt caatggagcc cagaagttgt
     3781 ttggaatgat cattactatc ggtcagtcca ttgtgtatgt gatgacgggc atgtacgggg
     3841 acccttcgga gatgggtgct ggaatctgcc tgctgatcac cattcagctc tttgttgctg
     3901 gcttaattgt cctacttttg gatgaacttc tgcaaaaagg gtacggcttg ggctctggca
     3961 tttctctctt cattgcgact aacatctgtg agaccattgt gtggaaggca ttcagcccca
     4021 ccactgtcaa cactggccga gggatggaat tcgaaggcgc catcatagcg ctattccacc
     4081 tgctggccac ccgcacagac aaggtccgag cccttcgaga ggcattctac cgccagaacc
     4141 tccccaacct catgaacctg attgccacca tcttcgtctt tgccgttgtc atctatttcc
     4201 agggcttccg agtggacctt ccaatcaagt cggcgcgtta ccgaggccag tacaacacct
     4261 atcccatcaa gctcttctac acctccaaca tccccatcat cttgcagtct gctctggtgt
     4321 ccaacctgta tgtcatctcc cagatgctgt cagcacgctt cagtggcaac ctgctggtta
     4381 gcctgctggg cacctggtcg gatacatcat ctgggggccc agcccgagct taccccgttg
     4441 gtggcctgtg ttactacctg tctcctccgg aatcgttcgg ctcagtgctg gaggaccctg
     4501 tccatgcggt cgtgtacatc gtgttcatgc tgggctcctg tgcattcttc tccaaaacat
     4561 ggatcgaagt ctcaggttct tctgccaaag atgttgcaaa gcagctgaag gagcagcaga
     4621 tggtgatgag gggccacagg gagacctcca tggtgcatga gctcaaccgg tacatcccca
     4681 cagccgcagc ctttggtggg ctgtgcattg gggccctctc tgtcctggct gacttcctgg
     4741 gtgctatcgg gtctggaact gggatcctgc ttgcggtcac catcatctac cagtactttg
     4801 agatcttcgt caaggaacag agcgaggtgg gcagcatggg agcccttctc ttctgagaat
     4861 tcctcgaggg cggccgctct agagtcgacg ggccgcggta acaattgtta actaacttaa
     4921 gctagcaacg gtttccctct agcgggatca attccgcccc ccccccctaa cgttactggc
     4981 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     5041 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     5101 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     5161 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     5221 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     5281 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     5341 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     5401 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     5461 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgataatac
     5521 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     5581 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     5641 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     5701 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     5761 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     5821 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     5881 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     5941 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     6001 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     6061 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     6121 acccgggcgg ggcgcgtctg gaacaatcaa cctctggatt acaaaatttg tgaaagattg
     6181 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     6241 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     6301 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     6361 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     6421 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     6481 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
     6541 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     6601 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     6661 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     6721 gccgcctccc cgcctggaat taattctgca gtcgagacct agaaaaacat ggagcaatca
     6781 caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg
     6841 aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag
     6901 ctgtagatct tagccacttt ttaaaagaaa agaggggact ggaagggcta attcactccc
     6961 aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt
     7021 agcagaacta cacaccaggg ccaggggtca gatatccact gacctttgga tggtgctaca
     7081 agctagtacc agttgagcca gataaggtag aagaggccaa taaaggagag aacaccagct
     7141 tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagtg ttagagtgga
     7201 ggtttgacag ccgcctagca tttcatcacg tggcccgaga gctgcatccg gagtacttca
     7261 agaactgctg atatcgagct tgctacaagg gactttccgc tggggacttt ccagggaggc
     7321 gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa gcagctgctt
     7381 tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     7441 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     7501 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     7561 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     7621 aaatgaatat cagagagtga gaggccttga cattgctagc gtttaccgtc gacctctagc
     7681 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     7741 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     7801 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     7861 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     7921 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     7981 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     8041 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     8101 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     8161 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     8221 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     8281 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     8341 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     8401 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     8461 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     8521 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     8581 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     8641 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     8701 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     8761 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     8821 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     8881 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     8941 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     9001 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     9061 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     9121 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     9181 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     9241 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     9301 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     9361 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     9421 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     9481 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     9541 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     9601 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     9661 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     9721 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     9781 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     9841 aaagtgccac ctgacgtcga cggatcggga gatcaacttg tttattgcag cttataatgg
     9901 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     9961 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac
    10021 tcaagctaac caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta
    10081 cctgtggttt catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa
    10141 ttgtatttgt taaatatgta ctacaaactt agtagt
//
