LOCUS       V028635                10089 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028635
VERSION     V028635
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..4845
                     /label="SLC24A2(NM_020344)"
                     /note="SLC24A2(NM_020344)"
                     /gene="SLC24A2"
     promoter        4877..5376
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5397..5996
                     /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    6010..6598
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6805..7438
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7567..7583)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7591..7607
                     /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(7615..7645)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7660..7681
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7969..8557)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8728..9588)
                     /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(9589..9693)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9741..9875
                     /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 tcatggatct gcaacaaagc
     2881 accaccatca cttccctaga gaaatggtgt ttggatgagt cactgtctgg ctgcagaaga
     2941 cattatagtg tcaagaaaaa actgaagtta attcgagtct taggcctttt catgggtctg
     3001 gtagccatta gcactgtctc attttcaatc agtgcctttt ctgagacaga tacacagagc
     3061 acaggagagg ccagtgttgt aagtggccct agggtagcac agggttacca tcagagaact
     3121 ctcttagatt taaatgacaa gattctggat tatactccac agccacctct ttctaaggaa
     3181 ggcgagtctg agaatagtac agatcacgcc caaggagact acccgaaaga catcttttcc
     3241 cttgaggaga gaagaaaagg tgcgatcatt ctgcatgtca ttggaatgat ctacatgttc
     3301 atagccttag ccattgtctg tgatgagttc tttgttcctt ctttgactgt catcactgaa
     3361 aaactgggca tctctgatga tgtggctgga gccaccttca tggctgcagg agggtcagcc
     3421 ccagaacttt tcacatctct cataggggta tttatcgctc acagcaacgt tggcataggc
     3481 acaattgtag gttcagcagt attcaacatc ctctttgtta ttggcatgtg tgctctgttt
     3541 tctagagaaa tcttaaacct gacatggtgg ccgctctttc gagatgtgtc tttctacatt
     3601 gttgacttga tcatgctgat catatttttc ctggataatg tcatcatgtg gtgggaaagc
     3661 ttgcttctct taacagctta tttttgctat gtggttttca tgaaattcaa cgtccaagta
     3721 gaaaaatggg tgaagcaaat gataaaccgc aataaggtcg tcaaggtgac agcaccagaa
     3781 gcccaagcaa agccatctgc agccagggac aaggatgaac caactctacc ggctaagccg
     3841 cgtctccagc gaggtggaag ctctgcctcc ctccacaaca gtctcatgag gaatagcatc
     3901 ttccaactca tgatacacac ccttgaccca ctcgccgaag aacttggatc atatggaaaa
     3961 ctaaaatatt atgacacaat gactgaagaa gggaggttca gagaaaaggc ttcaattctc
     4021 cacaagatcg ccaagaagaa atgtcatgtg gatgagaacg agaggcagaa tggggctgcc
     4081 aaccacgtgg aaaaaattga gcttccaaac agcaccagca cagatgttga aatgacacca
     4141 tccagtgatg cttcagaacc tgtacaaaat ggaaatctct cccacaacat tgaaggtgca
     4201 gaagcccaga ccgctgatga ggaggaggac cagcctctca gccttgcctg gccttctgaa
     4261 acccgcaagc aagtcacgtt tctgattgtt ttccccatag tgtttcctct ctggattacg
     4321 ttacctgacg ttcgcaaacc ttcatcgagg aagttttttc ccatcacgtt ctttggctcc
     4381 attacctgga ttgcagtatt ctcttacttg atggtctggt gggcgcacca ggttggagag
     4441 acaattggca tcagtgaaga gattatgggc ctgaccatct tggctgctgg gacctccatc
     4501 cctgatctta tcaccagtgt catagtggcc cggaaggggt taggggacat ggctgtgtcc
     4561 agctctgttg gaagcaacat ttttgacatc actgtagggc tcccactgcc ctggctcctg
     4621 tacaccgtca ttcacagatt ccagccagtg gctgtcagca gcaatggcct tttctgtgcc
     4681 atcgtccttc tcttcatcat gctgctcttc gtcatcctct ctatcgccct ctgcaagtgg
     4741 cgaatgaaca aaatcctggg cttcatcatg tttggcctct actttgtgtt cctggtggtg
     4801 agcgttctcc tagaagacag aattcttaca tgccccgtct ccatctaggg atcccgcgac
     4861 tctagataat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct
     4921 ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc
     4981 acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct
     5041 actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga
     5101 caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg
     5161 aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct
     5221 cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg
     5281 ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc
     5341 gcgctgttct cctcttcctc atctccgggc ctttcgacct gcagcccaag cttaccatga
     5401 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
     5461 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
     5521 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     5581 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     5641 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     5701 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     5761 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     5821 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     5881 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     5941 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgaccgc
     6001 gtctggaaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     6061 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     6121 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     6181 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     6241 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     6301 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     6361 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     6421 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     6481 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     6541 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     6601 ggaattaatt ctgcagtcga gacctagaaa aacatggagc aatcacaagt agcaatacag
     6661 cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc
     6721 cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     6781 actttttaaa agaaaagagg ggactggaag ggctaattca ctcccaacga agacaagata
     6841 tccttgatct gtggatctac cacacacaag gctacttccc tgattagcag aactacacac
     6901 cagggccagg ggtcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg
     6961 agccagataa ggtagaagag gccaataaag gagagaacac cagcttgtta caccctgtga
     7021 gcctgcatgg gatggatgac ccggagagag aagtgttaga gtggaggttt gacagccgcc
     7081 tagcatttca tcacgtggcc cgagagctgc atccggagta cttcaagaac tgctgatatc
     7141 gagcttgcta caagggactt tccgctgggg actttccagg gaggcgtggc ctgggcggga
     7201 ctggggagtg gcgagccctc agatcctgca tataagcagc tgctttttgc ctgtactggg
     7261 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     7321 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     7381 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     7441 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     7501 agtgagaggc cttgacattg ctagcgtttt accgtcgacc tctagctaga gcttggcgta
     7561 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     7621 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     7681 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     7741 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     7801 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     7861 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     7921 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     7981 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     8041 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     8101 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     8161 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     8221 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     8281 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     8341 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     8401 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     8461 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     8521 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     8581 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     8641 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     8701 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     8761 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     8821 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     8881 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     8941 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     9001 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     9061 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     9121 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     9181 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     9241 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     9301 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     9361 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     9421 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     9481 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     9541 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     9601 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     9661 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     9721 cgtcgacgga tcgggagatc aacttgttta ttgcagctta taatggttac aaataaagca
     9781 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     9841 ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa
     9901 atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt
     9961 tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa
    10021 tatgtactac aaacttagta gtttttaaag aaattgtatt tgttaaatat gtactacaaa
    10081 cttagtagt
//
