LOCUS       V032997                 9617 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032997
VERSION     V032997
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..3018
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="cyan variant of GFP"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNG
                     IKAHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="CFP"
     CDS             3034..4536
                     /label="SLC7A11(NM_014331)"
                     /note="SLC7A11(NM_014331)"
                     /gene="SLC7A11"
     promoter        4587..4798
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4811..5079
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5106..5705
                     /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    5712..6300
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6374..6607
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6679..6800
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6819..6954
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6992..7008)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7016..7032
                     /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(7040..7070)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7085..7106
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7394..7982)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8153..9013)
                     /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(9014..9118)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9592..9608
                     /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 catggtgagc aagggcgagg agctgttcac cggggtggtg
     2341 cccatcctgg tcgagctgga cggcgacgta aacggccaca ggttcagcgt gtccggcgag
     2401 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2461 ctgcccgtgc cctggcccac cctcgtgacc accctgacct ggggcgtgca gtgcttcagc
     2521 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2581 gtccaggagc gtaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2641 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2701 gacggcaaca tcctggggca caagctggag tacaactaca tcagccacaa cgtctatatc
     2761 accgccgaca agcagaagaa cggcatcaag gcccacttca agatccgcca caacatcgag
     2821 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2881 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     2941 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3001 atggacgagc tgtacaaggg aggaggggga agcatggtca gaaagcctgt tgtgtccacc
     3061 atctccaaag gaggttacct gcagggaaat gttaacggga ggctgccttc cctgggcaac
     3121 aaggagccac ctgggcagga gaaagtgcag ctgaagagga aagtcacttt actgagggga
     3181 gtctccatta tcattggcac catcattgga gcaggaatct tcatctctcc taagggcgtg
     3241 ctccagaaca cgggcagcgt gggcatgtct ctgaccatct ggacggtgtg tggggtcctg
     3301 tcactatttg gagctttgtc ttatgctgaa ttgggaacaa ctataaagaa atctggaggt
     3361 cattacacat atattttgga agtctttggt ccattaccag cttttgtacg agtctgggtg
     3421 gaactcctca taatacgccc tgcagctact gctgtgatat ccctggcatt tggacgctac
     3481 attctggaac cattttttat tcaatgtgaa atccctgaac ttgcgatcaa gctcattaca
     3541 gctgtgggca taactgtagt gatggtccta aatagcatga gtgtcagctg gagcgcccgg
     3601 atccagattt tcttaacctt ttgcaagctc acagcaattc tgataattat agtccctgga
     3661 gttatgcagc taattaaagg tcaaacgcag aactttaaag acgccttttc aggaagagat
     3721 tcaagtatta cgcggttgcc actggctttt tattatggaa tgtatgcata tgctggctgg
     3781 ttttacctca actttgttac tgaagaagta gaaaaccctg aaaaaaccat tccccttgca
     3841 atatgtatat ccatggccat tgtcaccatt ggctatgtgc tgacaaatgt ggcctacttt
     3901 acgaccatta atgctgagga gctgctgctt tcaaatgcag tggcagtgac cttttctgag
     3961 cggctactgg gaaatttctc attagcagtt ccgatctttg ttgccctctc ctgctttggc
     4021 tccatgaacg gtggtgtgtt tgctgtctcc aggttattct atgttgcgtc tcgagagggt
     4081 caccttccag aaatcctctc catgattcat gtccgcaagc acactcctct accagctgtt
     4141 attgttttgc accctttgac aatgataatg ctcttctctg gagacctcga cagtcttttg
     4201 aatttcctca gttttgccag gtggcttttt attgggctgg cagttgctgg gctgatttat
     4261 cttcgataca aatgcccaga tatgcatcgt cctttcaagg tgccactgtt catcccagct
     4321 ttgttttcct tcacatgcct cttcatggtt gccctttccc tctattcgga cccatttagt
     4381 acagggattg gcttcgtcat cactctgact ggagtccctg cgtattatct ctttattata
     4441 tgggacaaga aacccaggtg gtttagaata atgtcggaga aaataaccag aacattacaa
     4501 ataatactgg aagttgtacc agaagaagat aagttatagg gatccgcggc cgcgaaggat
     4561 ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa
     4621 gttgggggga ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg
     4681 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
     4741 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct
     4801 gaagcttcga ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca
     4861 tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc
     4921 cgccgtctag gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg
     4981 gagcctacct agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta
     5041 cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac
     5101 gctagatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg
     5161 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc
     5221 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     5281 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     5341 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     5401 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     5461 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     5521 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     5581 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     5641 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     5701 cctgagtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     5761 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     5821 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     5881 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     5941 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     6001 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     6061 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     6121 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     6181 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     6241 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     6301 ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     6361 gaaaaggggg gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct
     6421 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     6481 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     6541 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     6601 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     6661 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     6721 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6781 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
     6841 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6901 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact
     6961 tttgcagaga cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     7021 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     7081 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     7141 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     7201 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     7261 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     7321 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     7381 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     7441 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7501 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7561 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7621 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7681 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7741 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7801 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     7861 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7921 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7981 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     8041 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     8101 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     8161 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     8221 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     8281 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     8341 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     8401 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     8461 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     8521 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     8581 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     8641 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     8701 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8761 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8821 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8881 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8941 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     9001 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     9061 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     9121 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     9181 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     9241 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     9301 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     9361 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     9421 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     9481 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     9541 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     9601 cggccagtgc caagctg
//
