LOCUS       V020285                 8927 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020285
VERSION     V020285
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"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2826..3683
                     /label="Hoxb13(NM_001107041)"
                     /note="Hoxb13(NM_001107041)"
                     /gene="Hoxb13"
     promoter        3715..4214
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4235..4834
                     /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    4848..5436
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5643..6276
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6405..6421)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6429..6445
                     /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(6453..6483)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6498..6519
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6807..7395)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7566..8426)
                     /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(8427..8531)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8579..8713
                     /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 cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag
     2821 ccaccatgga gcccggcaat tatgccacct tggacggggc caaggatatc gaaggcttgc
     2881 tgggagctgg agggggtcgg aatttagtcg cccactcctc cccactggct agccatcccg
     2941 cagctccaac gctgatgccg actgtcaact atgccccctt ggatctgcca ggctctgcag
     3001 agccaccaaa gcagtgccac ccttgtcctg gggtgcccca gggggcatct ccagctcctg
     3061 tgccttatgg ctactttgga ggcgggtact actcttgccg agtatccagg agctcactga
     3121 aaccctgtgc ccagacggcc accctggcta cttacccttc ggaaacccct gcacctgggg
     3181 aggagtatcc tagccgtccc accgagtttg ccttctatcc gggctacccg ggaccttacc
     3241 agcctatggc cagttacctg gatgtgtctg tggtgcagac cctgggggcc cctggggagc
     3301 ctcgccacga ctctctgctt cccgtggaca gttatcagcc ttgggccctg gccggtggct
     3361 ggaacagcca gatgtgctgc caaggtgaac agaacccacc aggtccattc tggaaagcag
     3421 catttgcaga gcccagtgtc cagcaccctc ctcccgacgg ctgcgccttc cgccgcggcc
     3481 gcaaaaaacg cattccctat agcaaggggc agttgcgaga gctggagcga gagtatgcgg
     3541 ccaacaagtt tatcaccaag gacaagaggc gcaagatctc ggcagccacc agcctctctg
     3601 aacgccagat taccatctgg tttcagaacc gccgggtcaa ggagaagaag gttcttgcca
     3661 aggtcaagac cagcactact ccgtgaggat cccgcgactc tagataattc taccgggtag
     3721 gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt
     3781 ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac
     3841 cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt
     3901 tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca
     3961 ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca
     4021 gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt
     4081 ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag
     4141 gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat
     4201 ctccgggcct ttcgacctgc agcccaagct taccatgacc gagtacaagc ccacggtgcg
     4261 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     4321 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     4381 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     4441 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     4501 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     4561 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     4621 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     4681 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     4741 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     4801 cacctggtgc atgacccgca agcccggtgc ctgaccgcgt ctggaacaat caacctctgg
     4861 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     4921 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     4981 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     5041 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     5101 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     5161 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     5221 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     5281 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     5341 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     5401 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aattaattct gcagtcgaga
     5461 cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc
     5521 ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt
     5581 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagagggg
     5641 actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca
     5701 cacacaaggc tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc
     5761 actgaccttt ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc
     5821 caataaagga gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc
     5881 ggagagagaa gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg
     5941 agagctgcat ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc
     6001 cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag
     6061 atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct
     6121 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     6181 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     6241 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     6301 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggcct tgacattgct
     6361 agcgttttac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     6421 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6481 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6541 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6601 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6661 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6721 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6781 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6841 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6901 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6961 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7021 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7081 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7141 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7201 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     7261 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7321 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7381 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7441 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7501 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7561 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7621 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7681 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7741 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7801 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7861 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7921 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7981 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8041 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8101 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8161 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8221 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8281 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8341 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8401 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8461 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8521 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatcaa
     8581 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     8641 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     8701 tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc
     8761 ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct gtgattcctc
     8821 tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt
     8881 ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagt
//
