LOCUS       V046584                 9396 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046584
VERSION     V046584
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..1921
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2011..2214
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2303..3448
                     /label="Gpr132(NM_019925)"
                     /note="Gpr132(NM_019925)"
                     /gene="Gpr132"
     CDS             3455..3544
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        3594..3805
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3818..4086
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4121..4828
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             4838..4891
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             4892..5491
                     /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    5492..6079
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6153..6386
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6458..6579
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6598..6733
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6771..6787)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6795..6811
                     /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(6819..6849)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6864..6885
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7173..7761)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7932..8792)
                     /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(8793..8897)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9371..9387
                     /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 attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc gaattcgcca ccatgagatc agaacctacc aatgcagcag gaaacaccac
     2341 actgggggtt acctccgttc ttcagagcac ctcagtacct tcttctgaga cctgccacgt
     2401 ctcctacgag gagagcagag tggtcctggt ggtggtgtac agtgccgtgt gcctgctggg
     2461 cctaccagcc aactgcctga ctgcctggct gacgctgctg caagtcctgc agaggaacgt
     2521 gctagccgtc tacctgttct gcctgtccct ctgtgagctg ctctacatca gcacggtgcc
     2581 attgtggatc atctacatcc agaatcagca caaatggaac ctgggtccgc aggcctgcaa
     2641 ggtgactgct tacatcttct tctgcaacat ctacatcagc atcctcttgc tctgctgcat
     2701 ttcctgcgac cgctacatgg ccgtggtcta tgcactggag agccgaggcc accgccacca
     2761 gaggactgct gtcaccattt ctgcgtgtgt gattcttctt gttggacttg ttaactatcc
     2821 agtgtttgac atgaaggtgg agaagagttt ctgctttgag cccctgagga tgaacagcaa
     2881 gatagccggc taccactacc tgcgtttcac ctttggcttt gccatccctc tcggcatcct
     2941 ggcgttcacc aatcaccaga tcttccggag catcaaactc agtgacagcc tgagcgctgc
     3001 gcagaagaac aaggtgaagc gctccgccat cgcggtcgtc accatcttcc tggtctgctt
     3061 tgctccctac cacgtggtac tcctcgtcaa agctgccagc ttttccttct accaaggaga
     3121 catggatgcc gtgtgtgcct ttgaaagcag actgtacaca gtctctatgg tgtttctgtg
     3181 cctgtctaca gtcaacagtg tggctgaccc catcatctac gtgctgggta cagaccactc
     3241 tcggcaagaa gtgtccagaa tccacacagg gtggaaaaag tggtccacaa agacatatgt
     3301 tacatgctca aaggactctg aggagacaca cttgcccaca gagctttcaa acacatacac
     3361 cttccccaat cccgcgcacc ctccaggatc acagccagcg aagctaggtt tactgtgctc
     3421 gccagagaga ctgcctgagg agctctgcct cgaggaacaa aaactaatat cagaggaaga
     3481 tttggaacag aaactaatca gtgaagaaga tttagagcaa aagttaattt ccgaggagga
     3541 cttataggat ccgcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga
     3601 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc
     3661 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     3721 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     3781 caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg
     3841 cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc
     3901 cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag
     3961 accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct
     4021 ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta
     4081 cagatccaag ctgtgaccgg cgcctacgct agacgccacc atggtgagca agggcgagga
     4141 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
     4201 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     4261 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
     4321 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
     4381 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     4441 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
     4501 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
     4561 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
     4621 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
     4681 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
     4741 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
     4801 caccggcggc atggacgagc tgtacaaggg atctcgcgag ggcagaggaa gtcttctaac
     4861 atgcggtgac gtggaggaga atcccggccc tatgaccgag tacaagccca cggtgcgcct
     4921 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     4981 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     5041 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     5101 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     5161 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     5221 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     5281 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     5341 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     5401 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     5461 ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct ctggattaca aaatttgtga
     5521 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     5581 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     5641 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     5701 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     5761 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     5821 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     5881 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     5941 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     6001 gctgccggct ctgcggcctc ttccgcgtct ccgccttcgc cctcagacga gtcggatctc
     6061 cctttggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga
     6121 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaaa
     6181 ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     6241 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     6301 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     6361 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     6421 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     6481 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6541 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     6601 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     6661 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     6721 ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg
     6781 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6841 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6901 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6961 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     7021 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     7081 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     7141 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     7201 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7261 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7321 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7381 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7441 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7501 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7561 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     7621 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7681 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7741 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7801 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7861 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7921 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7981 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     8041 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     8101 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     8161 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     8221 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     8281 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     8341 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8401 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     8461 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8521 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8581 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8641 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8701 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8761 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8821 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8881 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     8941 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     9001 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     9061 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     9121 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     9181 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     9241 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     9301 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     9361 tcacgacgtt gtaaaacgac ggccagtgcc aagctg
//
