LOCUS       V039617                 9151 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039617
VERSION     V039617
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..3804
                     /label="SLC7A11(NM_014331)"
                     /note="SLC7A11(NM_014331)"
                     /gene="SLC7A11"
     CDS             3811..3876
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3920..4131
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4144..4412
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4445..5239
                     /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    5246..5834
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5908..6141
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6213..6334
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6353..6488
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6526..6542)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6550..6566
                     /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(6574..6604)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6619..6640
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6928..7516)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7687..8547)
                     /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(8548..8652)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9126..9142
                     /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 catggtcaga aagcctgttg tgtccaccat ctccaaagga
     2341 ggttacctgc agggaaatgt taacgggagg ctgccttccc tgggcaacaa ggagccacct
     2401 gggcaggaga aagtgcagct gaagaggaaa gtcactttac tgaggggagt ctccattatc
     2461 attggcacca tcattggagc aggaatcttc atctctccta agggcgtgct ccagaacacg
     2521 ggcagcgtgg gcatgtctct gaccatctgg acggtgtgtg gggtcctgtc actatttgga
     2581 gctttgtctt atgctgaatt gggaacaact ataaagaaat ctggaggtca ttacacatat
     2641 attttggaag tctttggtcc attaccagct tttgtacgag tctgggtgga actcctcata
     2701 atacgccctg cagctactgc tgtgatatcc ctggcatttg gacgctacat tctggaacca
     2761 ttttttattc aatgtgaaat ccctgaactt gcgatcaagc tcattacagc tgtgggcata
     2821 actgtagtga tggtcctaaa tagcatgagt gtcagctgga gcgcccggat ccagattttc
     2881 ttaacctttt gcaagctcac agcaattctg ataattatag tccctggagt tatgcagcta
     2941 attaaaggtc aaacgcagaa ctttaaagac gccttttcag gaagagattc aagtattacg
     3001 cggttgccac tggcttttta ttatggaatg tatgcatatg ctggctggtt ttacctcaac
     3061 tttgttactg aagaagtaga aaaccctgaa aaaaccattc cccttgcaat atgtatatcc
     3121 atggccattg tcaccattgg ctatgtgctg acaaatgtgg cctactttac gaccattaat
     3181 gctgaggagc tgctgctttc aaatgcagtg gcagtgacct tttctgagcg gctactggga
     3241 aatttctcat tagcagttcc gatctttgtt gccctctcct gctttggctc catgaacggt
     3301 ggtgtgtttg ctgtctccag gttattctat gttgcgtctc gagagggtca ccttccagaa
     3361 atcctctcca tgattcatgt ccgcaagcac actcctctac cagctgttat tgttttgcac
     3421 cctttgacaa tgataatgct cttctctgga gacctcgaca gtcttttgaa tttcctcagt
     3481 tttgccaggt ggctttttat tgggctggca gttgctgggc tgatttatct tcgatacaaa
     3541 tgcccagata tgcatcgtcc tttcaaggtg ccactgttca tcccagcttt gttttccttc
     3601 acatgcctct tcatggttgc cctttccctc tattcggacc catttagtac agggattggc
     3661 ttcgtcatca ctctgactgg agtccctgcg tattatctct ttattatatg ggacaagaaa
     3721 cccaggtggt ttagaataat gtcagagaaa ataaccagaa cattacaaat aatactggaa
     3781 gttgtaccag aagaagataa gttaggatcc gactacaaag accatgacgg tgattataaa
     3841 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat
     3901 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
     3961 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
     4021 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
     4081 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt
     4141 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc
     4201 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc
     4261 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta
     4321 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt
     4381 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctaggc
     4441 caccatggct gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4501 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4561 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4621 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4681 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4741 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4801 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4861 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4921 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     4981 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5041 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     5101 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     5161 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5221 tcttgacgag ttcttctgag tcgacaatca acctctggat tacaaaattt gtgaaagatt
     5281 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     5341 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     5401 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     5461 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     5521 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     5581 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     5641 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     5701 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     5761 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     5821 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta
     5881 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag
     5941 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     6001 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     6061 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     6121 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     6181 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     6241 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6301 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     6361 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     6421 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     6481 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc
     6541 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6601 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6661 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6721 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6781 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6841 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6901 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6961 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7021 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7081 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7141 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7201 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7261 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7321 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7381 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7441 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7501 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7561 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7621 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7681 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7741 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7801 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7861 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7921 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7981 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8041 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8101 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8161 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8221 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8281 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8341 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8401 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8461 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8521 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8581 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8641 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     8701 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     8761 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     8821 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     8881 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     8941 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     9001 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     9061 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     9121 acgttgtaaa acgacggcca gtgccaagct g
//
