LOCUS       V031616                10240 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031616
VERSION     V031616
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..5070
                     /label="CCAR2(NM_021174)"
                     /note="CCAR2(NM_021174)"
                     /gene="CCAR2"
     CDS             5077..5166
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        5210..5421
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5434..5702
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5729..6328
                     /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    6335..6923
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6997..7230
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7302..7423
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7442..7577
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7615..7631)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7639..7655
                     /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(7663..7693)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7708..7729
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8017..8605)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8776..9636)
                     /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(9637..9741)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     10215..10231
                     /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 catgtcccag tttaagcgcc agcggatcaa cccgcttcca
     2341 gggggacgca acttctcagg cacagcttca acatctcttc tgggccctcc tcctggtttg
     2401 ctcactcctc ctgtggccac agaactgtcc cagaatgcca ggcaccttca gggtggggag
     2461 aaacagcggg tcttcactgg tattgttacc agcttgcatg actactttgg ggttgtggat
     2521 gaagaggtct tttttcagct aagtgtggtg aagggccgtc tgccccagct gggtgagaag
     2581 gtgctggtga aggctgcata caacccaggc caggcagtgc cctggaatgc tgtcaaggtg
     2641 caaacgctct ccaaccagcc cctactgaag tccccagcac ctcctcttct gcatgtagca
     2701 gccctgggcc agaagcaagg gatcctggga gctcagcctc agttgatctt ccagcctcac
     2761 cggattcccc cactctttcc tcagaagcct ctgagtctct tccaaacatc ccacacactt
     2821 cacctgagcc acctgaacag atttcctgcc cggggccctc atggacggtt ggatcagggc
     2881 cgaagtgatg actatgactc caagaaacgc aaacagcggg ctggtggaga gccctggggt
     2941 gctaagaagc caaggcatga cctgcctcct taccgggtcc acctcactcc ttacactgtg
     3001 gacagcccca tctgtgactt cctagaactc cagcgccgtt accgcagcct cctggtcccc
     3061 tcagattttc tgtccgtgca tctgagttgg ctatcagcct tccccctgag ccagcccttt
     3121 tccctccatc atccaagccg gatccaggtc tcttctgaaa aggaggcagc tccagacgct
     3181 ggtgctgagc ccatcactgc agacagtgac cccgcttata gttcgaaggt actgctgctc
     3241 tcttccccgg ggttggagga attgtatcgt tgttgcatgc tctttgtgga tgacatggct
     3301 gagccaaggg agacgccaga gcatcctctg aagcagatta agtttttgct gggcaggaaa
     3361 gaagaggagg cagtgctggt tgggggtgaa tggtctcctt ccctggatgg cctcgacccc
     3421 caggctgacc cgcaggtgct ggtgcgtacc gccatccgct gtgcgcaggc ccagactggc
     3481 attgatttga gcggctgtac caagtggtgg cgctttgccg agtttcagta cctgcagccg
     3541 ggaccccccc ggcggcttca gacagtggtg gtgtacctgc cggatgtctg gaccatcatg
     3601 cctactttgg aggagtggga ggccctgtgc cagcagaaag ctgcagaggc agctccccca
     3661 acccaggagg cacaagggga aacggagcct actgaacagg cacctgatgc cttggagcaa
     3721 gcagcagaca cttctagacg gaacgcagaa actccagagg ccaccacaca gcaggaaacg
     3781 gacactgatc tcccagaggc ccctccaccc cccctagaac ctgctgtcat cgcacgccct
     3841 ggctgtgtaa acctgtccct ccatgggatt gtggaggatc ggaggccaaa ggaaaggatc
     3901 tcttttgagg tgatggtgct ggccgagctg tttctggaga tgctccagag ggattttggc
     3961 tatagagttt ataagatgct actgagcctt cctgaaaagg tcgtgtcccc acctgaacct
     4021 gagaaggagg aggcggccaa ggaagaagcc accaaggagg aagaagccat caaagaggag
     4081 gtggtcaagg agcccaagga tgaggcacag aatgagggcc cggctacaga gtcagaggcc
     4141 ccgctgaagg aggatgggct tttgcccaaa ccactctctt ctgggggaga ggaagaagaa
     4201 aaaccccggg gcgaggcttc tgaggacctg tgtgagatgg ccctggaccc agaactgttg
     4261 cttctgaggg atgatggaga ggaggagttt gcaggagcaa agctggagga ttcggaggtc
     4321 cggtccgttg cctcaaacca gtcagagatg gagttctctt cacttcagga catgcccaag
     4381 gagctggatc cctctgctgt gctcccctta gactgtctgc ttgcttttgt gttctttgat
     4441 gccaactggt gtggctactt gcaccggcga gacttagaga ggatcctcct tacccttggg
     4501 atccggctca gtgcagagca ggccaagcag ctggtcagca gggtggtgac ccagaacatc
     4561 tgccagtacc ggagccttca gtacagccgc caggagggcc tggatggtgg ccttcccgag
     4621 gaggtgctct tcggaaacct ggacctgctg ccccctcctg ggaaaagcac gaagccaggt
     4681 gctgccccca cagaacacaa agccttggtg tcccacaatg gcagcctgat taacgtgggg
     4741 agcctgctgc agcgcgcgga gcagcaggac agcggccggc tctacctaga gaacaagatc
     4801 cacacactgg agctgaagct ggaggagagc cataaccgtt tctcagccac tgaagtaacc
     4861 aataagacgc tggcggcaga gatgcaggag ctgcgagtcc ggctggcgga ggccgaggag
     4921 accgcccgga cggcggagcg acagaagagc cagctccagc ggctgctgca ggagctccgc
     4981 aggcgtctga cccccctgca gctggagatc cagcgggtgg tggaaaaggc tgacagctgg
     5041 gtggagaagg aggagccggc acctagcaac ggatccgaac aaaaactaat atcagaggaa
     5101 gatttggaac agaaactaat cagtgaagaa gatttagagc aaaagttaat ttccgaggag
     5161 gacttatagc ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc
     5221 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag
     5281 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     5341 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
     5401 gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc
     5461 cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc
     5521 tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg
     5581 ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc
     5641 ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga
     5701 tccaagctgt gaccggcgcc tacgctagat gaccgagtac aagcccacgg tgcgcctcgc
     5761 cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     5821 cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg agctgcaaga
     5881 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     5941 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     6001 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     6061 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     6121 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     6181 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     6241 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     6301 gtgcatgacc cgcaagcccg gtgcctgagt cgacaatcaa cctctggatt acaaaatttg
     6361 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     6421 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     6481 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     6541 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     6601 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     6661 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     6721 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     6781 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     6841 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     6901 ctccctttgg gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg
     6961 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     7021 gaaaataaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     7081 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     7141 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     7201 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     7261 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     7321 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     7381 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     7441 tatcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     7501 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     7561 aggctttttt ggaggcctag acttttgcag agacggccca aattcgtaat catggtcata
     7621 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     7681 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     7741 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     7801 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     7861 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7921 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7981 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     8041 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     8101 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     8161 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     8221 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     8281 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     8341 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     8401 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     8461 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     8521 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     8581 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     8641 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     8701 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     8761 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     8821 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     8881 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     8941 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     9001 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     9061 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     9121 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     9181 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     9241 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     9301 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     9361 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     9421 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     9481 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     9541 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     9601 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     9661 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     9721 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     9781 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     9841 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     9901 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     9961 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
    10021 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
    10081 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
    10141 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
    10201 ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
