LOCUS       V032575                 8712 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032575
VERSION     V032575
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    1798..1915
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2005..2208
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2297..2731
                     /label="SYNJ2BP(NM_018373)"
                     /note="SYNJ2BP(NM_018373)"
                     /gene="SYNJ2BP"
     CDS             2738..2827
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        2871..3082
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3095..3363
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3392..3401
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3422..4084
                     /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             4154..4207
                     /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             4208..4807
                     /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    4808..5395
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5469..5702
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5774..5895
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5914..6049
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6087..6103)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6111..6127
                     /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(6135..6165)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6180..6201
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6489..7077)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7248..8108)
                     /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(8109..8213)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8687..8703
                     /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 ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagcgaattc gccaccatga acggaagagt ggattatttg gtcactgagg aagagatcaa
     2341 tcttaccaga gggccctcag ggctgggctt caacatcgtc ggtgggacag atcagcagta
     2401 tgtctccaac gacagtggca tctacgtcag ccgcatcaaa gaaaatgggg ctgcggccct
     2461 ggatgggcgg ctccaggagg gtgataagat cctttcggta aatggccaag acctaaagaa
     2521 cctgctgcac caggatgctg tagacctctt tcgtaatgca ggctatgctg tgtctctgag
     2581 agtgcagcac aggttacagg tgcagaatgg acctatagga catcgaggtg aaggggaccc
     2641 aagtggtatt cccatattta tggtgctggt gccagtgttt gccctcacca tggtagcagc
     2701 ctgggctttc atgagatacc ggcaacaact tggatccgaa caaaaactaa tatcagagga
     2761 agatttggaa cagaaactaa tcagtgaaga agatttagag caaaagttaa tttccgagga
     2821 ggacttatag cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg
     2881 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta
     2941 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
     3001 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
     3061 cgggtttgcc gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc
     3121 ccgccgccct acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc
     3181 ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc
     3241 gggcctttgt ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg
     3301 cctgaccctg cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag
     3361 atccaagctg tgaccggcgc ctacgctaga cgccaccatg gagagcgacg agagcggcct
     3421 gcccgccatg gagatcgagt gccgcatcac cggcaccctg aacggcgtgg agttcgagct
     3481 ggtgggcggc ggagagggca cccccaagca gggccgcatg accaacaaga tgaagagcac
     3541 caaaggcgcc ctgaccttca gcccctacct gctgagccac gtgatgggct acggcttcta
     3601 ccacttcggc acctacccca gcggctacga gaaccccttc ctgcacgcca tcaacaacgg
     3661 cggctacacc aacacccgca tcgagaagta cgaggacggc ggcgtgctgc acgtgagctt
     3721 cagctaccgc tacgaggccg gccgcgtgat cggcgacttc aaggtggtgg gcaccggctt
     3781 ccccgaggac agcgtgatct tcaccgacaa gatcatccgc agcaacgcca ccgtggagca
     3841 cctgcacccc atgggcgata acgtgctggt gggcagcttc gcccgcacct tcagcctgcg
     3901 cgacggcggc tactacagct tcgtggtgga cagccacatg cacttcaaga gcgccatcca
     3961 ccccagcatc ctgcagaacg ggggccccat gttcgccttc cgccgcgtgg aggagctgca
     4021 cagcaacacc gagctgggca tcgtggagta ccagcacgcc ttcaagaccc ccatcgcctt
     4081 cgccagatcc cgcgctcagt cgtccaattc tgccgtggac ggcaccgccg gacccggctc
     4141 caccggatct cgcgagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc
     4201 cggccctatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     4261 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     4321 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     4381 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     4441 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     4501 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     4561 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     4621 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     4681 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     4741 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     4801 tgcctgaaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4861 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4921 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4981 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     5041 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     5101 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     5161 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     5221 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     5281 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     5341 gcgtctccgc cttcgccctc agacgagtcg gatctccctt tggccgcctc cccgcctggt
     5401 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     5461 ggggggactg gaagggctaa ttcactccca acgaaaataa gatctgcttt ttgcttgtac
     5521 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     5581 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     5641 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     5701 cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc
     5761 agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5821 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5881 catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccagttcc
     5941 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     6001 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc
     6061 agagacggcc caaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     6121 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     6181 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6241 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6301 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     6361 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6421 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6481 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6541 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6601 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6661 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6721 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6781 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6841 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6901 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     6961 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7021 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7081 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7141 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7201 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7261 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7321 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7381 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7441 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7501 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7561 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7621 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7681 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7741 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7801 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7861 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7921 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7981 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     8041 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     8101 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     8161 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     8221 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     8281 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     8341 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     8401 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     8461 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     8521 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
     8581 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     8641 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     8701 agtgccaagc tg
//
