LOCUS       V046412                 9548 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046412
VERSION     V046412
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..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..2994
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="red fluorescent protein from Entacmaea
                     quadricolor"
                     /transl_table=1
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
                     /label="TurboRFP"
     CDS             3031..4239
                     /label="PTEN(NM_000314)"
                     /note="PTEN(NM_000314)"
                     /gene="PTEN"
     CDS             4246..4272
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        4317..4528
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4541..4809
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4842..5636
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     misc_feature    5643..6231
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6305..6538
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6610..6731
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6750..6885
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6923..6939)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6947..6963
                     /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(6971..7001)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7016..7037
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7325..7913)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8084..8944)
                     /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(8945..9049)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9523..9539
                     /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 attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catgagcgag ctgatcaagg agaacatgca catgaagctg
     2341 tacatggagg gcaccgtgaa caaccaccac ttcaagtgca catccgaggg cgaaggcaag
     2401 ccctacgagg gcacccagac catgaagatc aaggtggtcg agggcggccc tctccccttc
     2461 gccttcgaca tcctggctac cagcttcatg tacggcagca aagccttcat caaccacacc
     2521 cagggcatcc ccgacttctt taagcagtcc ttccctgagg gcttcacatg ggagagaatc
     2581 accacatacg aagacggggg cgtgctgacc gctacccagg acaccagctt ccagaacggc
     2641 tgcatcatct acaacgtcaa gatcaacggg gtgaacttcc catccaacgg ccctgtgatg
     2701 cagaagaaaa cacgcggctg ggaggccaac accgagatgc tgtaccccgc tgacggcggc
     2761 ctgagaggcc acagccagat ggccctgaag ctcgtgggcg ggggctacct gcactgctcc
     2821 ttcaagacca catacagatc caagaaaccc gctaagaacc tcaagatgcc cggcttccac
     2881 ttcgtggacc acagactgga aagaatcaag gaggccgaca aagagaccta cgtcgagcag
     2941 cacgagatgg ctgtggccaa gtactgcgac ctccctagca aactggggca cagactcgag
     3001 ggtggaggcg gatcaggagg cggtggatct atgacagcca tcatcaaaga gatcgttagc
     3061 agaaacaaaa ggagatatca agaggatgga ttcgacttag acttgaccta tatttatcca
     3121 aacattattg ctatgggatt tcctgcagaa agacttgaag gcgtatacag gaacaatatt
     3181 gatgatgtag taaggttttt ggattcaaag cataaaaacc attacaagat atacaatctt
     3241 tgtgctgaaa gacattatga caccgccaaa tttaattgca gagttgcaca atatcctttt
     3301 gaagaccata acccaccaca gctagaactt atcaaaccct tttgtgaaga tcttgaccaa
     3361 tggctaagtg aagatgacaa tcatgttgca gcaattcact gtaaagctgg aaagggacga
     3421 actggtgtaa tgatatgtgc atatttatta catcggggca aatttttaaa ggcacaagag
     3481 gccctagatt tctatgggga agtaaggacc agagacaaaa agggagtaac tattcccagt
     3541 cagaggcgct atgtgtatta ttatagctac ctgttaaaga atcatctgga ttatagacca
     3601 gtggcactgt tgtttcacaa gatgatgttt gaaactattc caatgttcag tggcggaact
     3661 tgcaatcctc agtttgtggt ctgccagcta aaggtgaaga tatattcctc caattcagga
     3721 cccacacgac gggaagacaa gttcatgtac tttgagttcc ctcagccgtt acctgtgtgt
     3781 ggtgatatca aagtagagtt cttccacaaa cagaacaaga tgctaaaaaa ggacaaaatg
     3841 tttcactttt gggtaaatac attcttcata ccaggaccag aggaaacctc agaaaaagta
     3901 gaaaatggaa gtctatgtga tcaagaaatc gatagcattt gcagtataga gcgtgcagat
     3961 aatgacaagg aatatctagt acttacttta acaaaaaatg atcttgacaa agcaaataaa
     4021 gacaaagcca accgatactt ttctccaaat tttaaggtga agctgtactt cacaaaaaca
     4081 gtagaggagc cgtcaaatcc agaggctagc agttcaactt ctgtaacacc agatgttagt
     4141 gacaatgaac ctgatcatta tagatattct gacaccactg actctgatcc agagaatgaa
     4201 ccttttgatg aagatcagca tacacaaatt acaaaagtcg gatcctaccc atacgatgtt
     4261 ccagattacg cttaggcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc
     4321 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg
     4381 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     4441 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     4501 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc
     4561 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc
     4621 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc
     4681 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac
     4741 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg
     4801 ttacagatcc aagctgtgac cggcgcctac gctaggccac catggctgaa caagatggat
     4861 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4921 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4981 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     5041 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5101 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5161 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5221 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5281 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5341 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     5401 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5461 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5521 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5581 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagtcg
     5641 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     5701 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     5761 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     5821 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     5881 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     5941 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     6001 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     6061 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     6121 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     6181 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct
     6241 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     6301 ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg
     6361 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     6421 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     6481 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     6541 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     6601 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6661 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6721 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc
     6781 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     6841 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag
     6901 acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6961 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     7021 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     7081 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     7141 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     7201 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     7261 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     7321 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     7381 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     7441 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     7501 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     7561 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     7621 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     7681 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7741 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     7801 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7861 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7921 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7981 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     8041 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     8101 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     8161 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     8221 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     8281 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     8341 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     8401 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     8461 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     8521 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     8581 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     8641 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     8701 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8761 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8821 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8881 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8941 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     9001 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     9061 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     9121 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     9181 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     9241 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     9301 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     9361 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     9421 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     9481 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     9541 ccaagctg
//
