LOCUS       V022848                 8855 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022848
VERSION     V022848
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             2317..2877
                     /label="SKAP2(NM_001303468)"
                     /note="SKAP2(NM_001303468)"
                     /gene="SKAP2"
     CDS             2884..2949
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3014..3225
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3238..3506
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3535..3544
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3565..4227
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     CDS             4297..4350
                     /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             4351..4950
                     /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    4951..5538
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5612..5845
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5917..6038
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6057..6192
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6230..6246)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6254..6270
                     /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(6278..6308)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6323..6344
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6632..7220)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7391..8251)
                     /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(8252..8356)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8830..8846
                     /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 ctgatacgaa ctcggaattc gccaccatga ataacactct aagaaaggat
     2341 ggaaagaaag attgctgttt tgaaatctct gctcctgata aacgtatata tcagtttaca
     2401 gcagcttctc ccaaagatgc tgaagaatgg gtacagcagc tgaaatttgt attgcaagat
     2461 atggaatctg atattattcc tgaggattat gatgagagag gagaattata tgatgatgtt
     2521 gatcatcctc taccaataag caatccacta acaagcagtc aaccaataga tgatgaaatt
     2581 tatgaagaac ttccagaaga agaagaggac agtgctccag tgaaagtgga agaacaaagg
     2641 aagatgagtc aggatagtgt ccatcacacc tcaggggata agagcactga ttatgctaat
     2701 ttttaccagg gattgtggga ttgtactgga gctttttctg atgagttgtc atttaagcgt
     2761 ggtgatgtga tttacattct tagcaaggaa tacaatagat atggctggtg ggtaggagaa
     2821 atgaagggag ccattggctt ggtgcctaaa gcctacataa tggagatgta tgatattctc
     2881 gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     2941 gatgacaagt agggatccct gcgatatcca gcgcggccgc gaaggatctg cgatcgctcc
     3001 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     3061 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     3121 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     3181 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg
     3241 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg
     3301 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta
     3361 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga
     3421 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg
     3481 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agacgccacc
     3541 atggagagcg acgagagcgg cctgcccgcc atggagatcg agtgccgcat caccggcacc
     3601 ctgaacggcg tggagttcga gctggtgggc ggcggagagg gcacccccaa gcagggccgc
     3661 atgaccaaca agatgaagag caccaaaggc gccctgacct tcagccccta cctgctgagc
     3721 cacgtgatgg gctacggctt ctaccacttc ggcacctacc ccagcggcta cgagaacccc
     3781 ttcctgcacg ccatcaacaa cggcggctac accaacaccc gcatcgagaa gtacgaggac
     3841 ggcggcgtgc tgcacgtgag cttcagctac cgctacgagg ccggccgcgt gatcggcgac
     3901 ttcaaggtgg tgggcaccgg cttccccgag gacagcgtga tcttcaccga caagatcatc
     3961 cgcagcaacg ccaccgtgga gcacctgcac cccatgggcg ataacgtgct ggtgggcagc
     4021 ttcgcccgca ccttcagcct gcgcgacggc ggctactaca gcttcgtggt ggacagccac
     4081 atgcacttca agagcgccat ccaccccagc atcctgcaga acgggggccc catgttcgcc
     4141 ttccgccgcg tggaggagct gcacagcaac accgagctgg gcatcgtgga gtaccagcac
     4201 gccttcaaga cccccatcgc cttcgccaga tcccgcgctc agtcgtccaa ttctgccgtg
     4261 gacggcaccg ccggacccgg ctccaccgga tctcgcgagg gcagaggaag tcttctaaca
     4321 tgcggtgacg tggaggagaa tcccggccct atgaccgagt acaagcccac ggtgcgcctc
     4381 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     4441 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     4501 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     4561 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     4621 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     4681 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     4741 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     4801 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     4861 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     4921 tggtgcatga cccgcaagcc cggtgcctga aatcaacctc tggattacaa aatttgtgaa
     4981 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     5041 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     5101 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     5161 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     5221 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     5281 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     5341 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     5401 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     5461 ctgccggctc tgcggcctct tccgcgtctc cgccttcgcc ctcagacgag tcggatctcc
     5521 ctttggccgc ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat
     5581 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaaa
     5641 taagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     5701 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     5761 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     5821 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     5881 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     5941 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6001 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     6061 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6121 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     6181 tttttggagg cctagacttt tgcagagacg gcccaaattc gtaatcatgg tcatagctgt
     6241 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6301 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6361 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6421 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6481 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6541 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6601 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6661 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6721 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6781 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6841 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6901 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6961 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7021 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     7081 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7141 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7201 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7261 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7321 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7381 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7441 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7501 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7561 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7621 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7681 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7741 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7801 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7861 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7921 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7981 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8041 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8101 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8161 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8221 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8281 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8341 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     8401 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     8461 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     8521 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     8581 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     8641 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
     8701 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     8761 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     8821 cacgacgttg taaaacgacg gccagtgcca agctg
//
