LOCUS       V026932                 9989 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026932
VERSION     V026932
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..4715
                     /label="SLC6A4(NM_001045)"
                     /note="SLC6A4(NM_001045)"
                     /gene="SLC6A4"
     CDS             4722..4745
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        4777..5276
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5297..5896
                     /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    5910..6498
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6705..7338
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7467..7483)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7491..7507
                     /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(7515..7545)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7560..7581
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7869..8457)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8628..9488)
                     /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(9489..9593)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9641..9775
                     /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 gacgacgccc ttgaattctc agaagcagct atcagcgtgt gaagatggag
     2881 aagattgtca ggaaaacgga gttctacaga aggttgttcc caccccaggg gacaaagtgg
     2941 agtccgggca aatatccaat gggtactcag cagttccaag tcctggtgcg ggagatgaca
     3001 cacggcactc tatcccagcg accaccacca ccctagtggc tgagcttcat caaggggaac
     3061 gggagacctg gggcaagaag gtggatttcc ttctctcagt gattggctat gctgtggacc
     3121 tgggcaatgt ctggcgcttc ccctacatat gttaccagaa tggagggggg gcattcctcc
     3181 tcccctacac catcatggcc atttttgggg gaatcccgct cttttacatg gagctcgcac
     3241 tgggacagta ccaccgaaat ggatgcattt caatatggag gaaaatctgc ccgattttca
     3301 aagggattgg ttatgccatc tgcatcattg ccttttacat tgcttcctac tacaacacca
     3361 tcatggcctg ggcgctatac tacctcatct cctccttcac ggaccagctg ccctggacca
     3421 gctgcaagaa ctcctggaac actggcaact gcaccaatta cttctccgag gacaacatca
     3481 cctggaccct ccattccacg tcccctgctg aagaatttta cacgcgccac gtcctgcaga
     3541 tccaccggtc taaggggctc caggacctgg ggggcatcag ctggcagctg gccctctgca
     3601 tcatgctgat cttcactgtt atctacttca gcatctggaa aggcgtcaag acctctggca
     3661 aggtggtgtg ggtgacagcc accttccctt atatcatcct ttctgtcctg ctggtgaggg
     3721 gtgccaccct ccctggagcc tggaggggtg ttctcttcta cttgaaaccc aattggcaga
     3781 aactcctgga gacaggggtg tggatagatg cagccgctca gatcttcttc tctcttggtc
     3841 cgggctttgg ggtcctgctg gcttttgcta gctacaacaa gttcaacaac aactgctacc
     3901 aagatgccct ggtgaccagc gtggtgaact gcatgacgag cttcgtttcg ggatttgtca
     3961 tcttcacagt gctcggttac atggctgaga tgaggaatga agatgtgtct gaggtggcca
     4021 aagacgcagg tcccagcctc ctcttcatca cgtatgcaga agcgatagcc aacatgccag
     4081 cgtccacttt ctttgccatc atcttctttc tgatgttaat cacgctgggc ttggacagca
     4141 cgtttgcagg cttggagggg gtgatcacgg ctgtgctgga tgagttccca cacgtctggg
     4201 ccaagcgccg ggagcggttc gtgctcgccg tggtcatcac ctgcttcttt ggatccctgg
     4261 tcaccctgac ttttggaggg gcctacgtgg tgaagctgct ggaggagtat gccacggggc
     4321 ccgcagtgct cactgtcgcg ctgatcgaag cagtcgctgt gtcttggttc tatggcatca
     4381 ctcagttctg cagggacgtg aaggaaatgc tcggcttcag cccggggtgg ttctggagga
     4441 tctgctgggt ggccatcagc cctctgtttc tcctgttcat catttgcagt tttctgatga
     4501 gcccgccaca actacgactt ttccaatata attatcctta ctggagtatc atcttgggtt
     4561 actgcatagg aacctcatct ttcatttgca tccccacata tatagcttat cggttgatca
     4621 tcactccagg gacatttaaa gagcgtatta ttaaaagtat taccccagaa acaccaacag
     4681 aaattccttg tggggacatc cgcttgaatg ctgtggaatt cgattacaag gatgacgacg
     4741 ataagtgagg atcccgcgac tctagataat tctaccgggt aggggaggcg cttttcccaa
     4801 ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc
     4861 tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg
     4921 gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc
     4981 tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg
     5041 acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt
     5101 gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg
     5161 gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc
     5221 ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct
     5281 gcagcccaag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     5341 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     5401 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     5461 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     5521 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     5581 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     5641 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     5701 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     5761 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     5821 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     5881 caagcccggt gcctgaccgc gtctggaaca atcaacctct ggattacaaa atttgtgaaa
     5941 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     6001 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     6061 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     6121 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     6181 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     6241 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     6301 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     6361 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     6421 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     6481 tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa aacatggagc
     6541 aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga
     6601 ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa
     6661 ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag ggctaattca
     6721 ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc
     6781 tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct ttggatggtg
     6841 ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag gagagaacac
     6901 cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtgttaga
     6961 gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc atccggagta
     7021 cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg actttccagg
     7081 gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc
     7141 tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     7201 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     7261 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     7321 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     7381 caaagaaatg aatatcagag agtgagaggc cttgacattg ctagcgtttt accgtcgacc
     7441 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     7501 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     7561 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     7621 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     7681 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     7741 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     7801 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     7861 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     7921 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     7981 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     8041 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     8101 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     8161 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     8221 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     8281 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     8341 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     8401 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     8461 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     8521 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     8581 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     8641 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     8701 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     8761 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     8821 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     8881 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     8941 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     9001 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     9061 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     9121 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     9181 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     9241 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     9301 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     9361 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     9421 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     9481 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     9541 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     9601 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc aacttgttta ttgcagctta
     9661 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     9721 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg
     9781 gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc
     9841 caattacctg tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta
     9901 aagaaattgt atttgttaaa tatgtactac aaacttagta gtttttaaag aaattgtatt
     9961 tgttaaatat gtactacaaa cttagtagt
//
