LOCUS       V028546                 9972 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028546
VERSION     V028546
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             2830..2856
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2863..4728
                     /label="SLC24A4(NM_153646)"
                     /note="SLC24A4(NM_153646)"
                     /gene="SLC24A4"
     promoter        4760..5259
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5280..5879
                     /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    5893..6481
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6688..7321
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7450..7466)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7474..7490
                     /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(7498..7528)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7543..7564
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7852..8440)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8611..9471)
                     /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(9472..9576)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9624..9758
                     /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 gccaccatgt acccatacga cgtaccagat tacgctgaat tcatggcgct ccgcgggacc
     2881 ctccggccgc tcaaagttcg caggaggcga gagatgctgc cgcagcaagt cggcttcgtg
     2941 tgcgcggtgc tggccctggt gtgctgtgcg tccggcctct tcggcagctt ggggcacaaa
     3001 acagcttctg ctagcaaacg tgtcctgcca gacacgtgga gaaatagaaa gttgatggcc
     3061 ccagtgaatg ggacacagac agccaagaac tgcacagatc ctgcgattca cgagttcccc
     3121 acagatctgt tctccaataa ggagcgacag cacggagccg tcctgctgca catccttggt
     3181 gctctgtata tgttctatgc cttggccata gtgtgcgatg acttctttgt tccgtctcta
     3241 gagaagatct gtgagagact ccatctgagc gaagatgtgg ctggagccac cttcatggct
     3301 gcaggaagct caacgccaga gctgtttgcg tctgttattg gggtgttcat cacccatggg
     3361 gacgtcgggg tgggcaccat cgtgggctct gctgtgttca acatcctgtg cataattgga
     3421 gtgtgcggac tgtttgctgg ccaggtggtc cgtctgacgt ggtgggccgt gtgccgagac
     3481 tccgtgtact acaccatctc tgtcatcgtg ctcatcgtgt tcatatatga tgaacaaatt
     3541 gtgtggtggg aaggcctggt gctcatcatc ttgtatgtgt tttatattct gatcatgaag
     3601 tacaatgtga agatgcaagc ctttttcaca gtcaaacaaa agagcattgc aaacggtaac
     3661 ccggtcaaca gtgagctgga ggctggtaat gatttctatg acggtagcta tgatgaccct
     3721 tccgtgccat tgctggggca agtgaaggag aagccacagt atggcaagaa ccccgtggtg
     3781 atggtggacg agattatgag ctccagccct cccaagttca ccttccctga agcaggctta
     3841 cgaatcatga tcaccaataa gtttggaccc aggacccgac tacggatggc cagcaggatc
     3901 atcattaatg agcggcagag actgatcaac tcggccaatg gtgtgagcag taagccgctt
     3961 caaaacggga ggcacgagaa cattgagaac gggaatgttc ctgtggaaaa ccccgaagac
     4021 cctcagcaga atcaggagca gcagccgccg ccacagccac caccgccaga gccagagccg
     4081 gtggaggctg acttcctgtc ccccttctcc gtgccggagg ccagagggga caaggtcaag
     4141 tgggtgttca cctggcccct catcttcctc ctgtgcgtca ccattcccaa ctgcagcaag
     4201 ccccgctggg agaagttctt catggtcacc ttcatcaccg ccacgctgtg gatcgctgtg
     4261 ttctcctaca tcatggtgtg gctggtgact attatcggat acacacttgg gatcccggat
     4321 gtcatcatgg gcattacttt cctggcagca gggacaagtg ttccagactg catggccagc
     4381 ctaattgtgg cgagacaagg ccttggggac atggcagtct ccaacaccat aggaagcaac
     4441 gtgtttgaca tcctggtagg acttggtgta ccgtggggcc tgcagaccat ggttgttaat
     4501 tatggatcaa cagtgaagat caacagccgg gggctggtct attccgtggt cctgttgctg
     4561 ggctctgtcg ctctcaccgt cctcggcatc cacctaaaca agtggcgact ggaccggaag
     4621 ctgggtgtct acgtgctggt tctctacgcc atcttcttgt gcttctccat aatgatagag
     4681 tttaacgtct ttaccttcgt caacttgccg atgtgccggg aagacgatta gggatcccgc
     4741 gactctagat aattctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc
     4801 gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat
     4861 tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct
     4921 tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg
     4981 tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa
     5041 tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg
     5101 gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg
     5161 gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct
     5221 gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagccc aagcttacca
     5281 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     5341 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     5401 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     5461 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     5521 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     5581 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     5641 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     5701 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     5761 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     5821 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac
     5881 cgcgtctgga acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     5941 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     6001 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     6061 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     6121 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     6181 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     6241 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     6301 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     6361 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     6421 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     6481 cctggaatta attctgcagt cgagacctag aaaaacatgg agcaatcaca agtagcaata
     6541 cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt
     6601 ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct gtagatctta
     6661 gccacttttt aaaagaaaag aggggactgg aagggctaat tcactcccaa cgaagacaag
     6721 atatccttga tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca
     6781 caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag
     6841 ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg
     6901 tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc
     6961 gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat
     7021 atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg
     7081 ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact
     7141 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     7201 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     7261 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     7321 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     7381 gagagtgaga ggccttgaca ttgctagcgt tttaccgtcg acctctagct agagcttggc
     7441 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     7501 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     7561 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     7621 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     7681 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     7741 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     7801 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     7861 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7921 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7981 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     8041 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     8101 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     8161 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     8221 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     8281 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     8341 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     8401 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     8461 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     8521 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     8581 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     8641 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     8701 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     8761 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8821 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8881 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8941 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     9001 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     9061 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     9121 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     9181 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     9241 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     9301 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     9361 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     9421 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     9481 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     9541 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     9601 tgacgtcgac ggatcgggag atcaacttgt ttattgcagc ttataatggt tacaaataaa
     9661 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     9721 tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact caagctaacc
     9781 aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac ctgtggtttc
     9841 atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat tgtatttgtt
     9901 aaatatgtac tacaaactta gtagttttta aagaaattgt atttgttaaa tatgtactac
     9961 aaacttagta gt
//
