LOCUS       V019095                 9630 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019095
VERSION     V019095
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..3807
                     /label="Fto(NM_011936)"
                     /note="Fto(NM_011936)"
                     /gene="Fto"
     promoter        3851..4062
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4075..4343
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4378..5070
                     /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             5071..5124
                     /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             5125..5724
                     /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    5725..6313
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6387..6620
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6692..6813
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6832..6967
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7005..7021)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7029..7045
                     /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(7053..7083)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7098..7119
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7407..7995)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8166..9026)
                     /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(9027..9131)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9605..9621
                     /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 catgaagcgc gtccagaccg cggaggaacg agagcgggaa
     2341 gctaagaaac tgaggctcct tgaggagctt gaagacactt ggcttcctta cctgaccccc
     2401 aaagatgatg agttctatca gcagtggcag ctgaaatacc ctaaactggt tttccgagag
     2461 gccggcagca taccagagga gctgcataag gaggtccccg aggcctttct cacactgcat
     2521 aagcatggct gcttgtttcg ggacgtggtg aggatccaag gcaaagatgt gctcacccca
     2581 gtgtctcgca tcctcatcgg ggacccaggc tgcacctaca agtacttgaa caccagactc
     2641 ttcacggtgc cctggcccgt gaagggctgc acggtcaagt acacagaggc tgagatcgcc
     2701 gctgcatgtc agaccttcct aaagctcaat gactacctcc aggtggagac catccaggcc
     2761 ttggaagaac tggctgtcag agagaaggcc aatgaagacg ctgtgccact gtgcatggca
     2821 gagttcccca gggccggcgt ggggccgtcc tgcgatgatg aagtggacct taagagcaga
     2881 gcagcctaca acgtgacttt gctaaacttc atggatcctc agaagatgcc ctacttgaaa
     2941 gaggagccct atttcggcat ggggaagatg gcggtgagct ggcatcacga tgagaacctg
     3001 gtggacaggt cagccgtggc agtgtacagc tatagctgcg aaggctctga ggatgaaagt
     3061 gaggacgagt ccagcttcga aggcagagat cctgatactt ggcatgttgg ttttaagatc
     3121 tcttgggaca tcgagacacc aggattaaca atccctcttc accagggaga ctgctatttc
     3181 atgctggatg acctcaatgc cacccaccag cactgtgttt tggctggctc acagcctcgg
     3241 tttagttcca ctcaccgtgt ggcagagtgc tcaacaggca ccttggatta tatcttagaa
     3301 cgctgtcagt tggcgctgca gaatgtcctc aatgactcag acgatggcga cgtctcgttg
     3361 aaatcctttg atcctgcagt tttgaaacaa ggagaggaaa tccataatga ggtggagttt
     3421 gagtggctga ggcagttctg gtttcaaggc aatcgataca aactttgcac cgattggtgg
     3481 tgtgagccca tgactcacct ggaggggctg tggaagaaga tggagagcat gacaaatgcg
     3541 gtgctccgtg aagttaaaag agaggggctc ccggtggaac aaaggagtga gattctgtct
     3601 gccatcctgg tcccgctcac cgtgcgccag aacctgagga aggagtggca tgccaggtgc
     3661 cagtcccgag tcgtccggac tttaccagta cagcagaaac cagactgccg gccatattgg
     3721 gagaaggatg acccttccat gcctctgccc tttgacctca cagacgtggt ttccgagctc
     3781 agaggccagc tgctggaagc aagatcctag gcggccgcaa ggatctgcga tcgctccggt
     3841 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     3901 ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     3961 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     4021 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct tcgaggggct
     4081 cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg ccggttgagt
     4141 cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt
     4201 ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct acctagactc
     4261 agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt tgtttcgttt
     4321 tctgttctgc gccgttacag atccaagctg tgaccggcgc ctacgctaga cgccaccatg
     4381 agcgagctga tcaaggagaa catgcacatg aagctgtaca tggagggcac cgtgaacaac
     4441 caccacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac ccagaccatg
     4501 aagatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct ggctaccagc
     4561 ttcatgtacg gcagcaaagc cttcatcaac cacacccagg gcatccccga cttctttaag
     4621 cagtccttcc ctgagggctt cacatgggag agaatcacca catacgaaga cgggggcgtg
     4681 ctgaccgcta cccaggacac cagcttccag aacggctgca tcatctacaa cgtcaagatc
     4741 aacggggtga acttcccatc caacggccct gtgatgcaga agaaaacacg cggctgggag
     4801 gccaacaccg agatgctgta ccccgctgac ggcggcctga gaggccacag ccagatggcc
     4861 ctgaagctcg tgggcggggg ctacctgcac tgctccttca agaccacata cagatccaag
     4921 aaacccgcta agaacctcaa gatgcccggc ttccacttcg tggaccacag actggaaaga
     4981 atcaaggagg ccgacaaaga gacctacgtc gagcagcacg agatggctgt ggccaagtac
     5041 tgcgacctcc ctagcaaact ggggcacaga gagggcagag gaagtcttct aacatgcggt
     5101 gacgtggagg agaatcccgg ccctatgacc gagtacaagc ccacggtgcg cctcgccacc
     5161 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     5221 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     5281 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     5341 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     5401 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     5461 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     5521 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     5581 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     5641 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     5701 atgacccgca agcccggtgc ctgaaatcaa cctctggatt acaaaatttg tgaaagattg
     5761 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     5821 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     5881 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     5941 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     6001 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     6061 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     6121 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     6181 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     6241 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     6301 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     6361 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaaaataaga
     6421 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     6481 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     6541 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     6601 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     6661 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     6721 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6781 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     6841 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     6901 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     6961 ggaggcctag acttttgcag agacggccca aattcgtaat catggtcata gctgtttcct
     7021 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     7081 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     7141 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     7201 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7261 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7321 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7381 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7441 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7501 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7561 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7621 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7681 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7741 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7801 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     7861 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7921 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7981 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     8041 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     8101 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     8161 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     8221 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     8281 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8341 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8401 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8461 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8521 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8581 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8641 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8701 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8761 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8821 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8881 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8941 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     9001 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     9061 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     9121 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     9181 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     9241 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     9301 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     9361 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     9421 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
     9481 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     9541 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     9601 cgttgtaaaa cgacggccag tgccaagctg
//
