LOCUS       V028438                 9363 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028438
VERSION     V028438
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        3..229
                     /note="Rous sarcoma virus enhancer/promoter"
                     /label="RSV promoter"
     LTR             230..410
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    457..582
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1075..1308
                     /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    1803..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     protein_bind    1938..1958
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of attB"
                     /note="core recombination site for the Gateway® BP
                     reaction"
                     /label="attB4"
     enhancer        2019..2322
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2323..2526
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2566..4032
                     /label="LMBR1L(NM_018113)"
                     /note="LMBR1L(NM_018113)"
                     /gene="LMBR1L"
     misc_feature    4078..4666
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     promoter        4697..5196
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5213..5812
                     /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"
     LTR             5884..6117
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6189..6310
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6350..6485
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(6506..6524)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(6534..6550)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      6692..7147
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        7173..7277
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             7278..8138
                     /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"
     rep_origin      8309..8897
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    9185..9206
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        9221..9251
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    9259..9275
                     /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"
     primer_bind     9283..9299
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        9320..9338
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /label="T3 promoter"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga 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 ggtatcgcta
     1801 gcttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
     1861 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt
     1921 actagtgatt atcggatcaa ctttgtatag aaaagttgta gttattaata gtaatcaatt
     1981 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     2041 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     2101 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     2161 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     2221 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     2281 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     2341 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     2401 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     2461 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     2521 agagctggtt tagtgaaccg tcagatccaa gttggatccg ccaccatgga agcacctgac
     2581 tacgaagtgc tatccgtgcg agaacagcta ttccacgaga ggatccgcga gtgtattata
     2641 tcaacacttc tgtttgcaac actgtacatc ctctgccaca tcttcctgac ccgcttcaag
     2701 aagcctgctg agttcaccac agtggatgat gaagatgcca ccgtcaacaa gattgcgctc
     2761 gagctgtgca cctttaccct ggcaattgcc ctgggtgctg tcctgctcct gcccttctcc
     2821 atcatcagca atgaggtgct gctctccctg cctcggaact actacatcca gtggctcaac
     2881 ggctccctca tccatggcct ctggaacctt gtttttctct tctccaacct gtccctcatc
     2941 ttcctcatgc cctttgcata tttcttcact gagtctgagg gctttgctgg ctccagaaag
     3001 ggtgtcctgg gccgggtcta tgagacagtg gtgatgttga tgctcctcac tctgctggtg
     3061 ctaggtatgg tgtgggtggc atcagccatt gtggacaaga acaaggccaa cagagagtca
     3121 ctctatgact tttgggagta ctatctcccc tacctctact catgcatctc cttccttggg
     3181 gttctgctgc tcctggtgtg tactccactg ggtctcgccc gcatgttctc cgtcactggg
     3241 aagctgctag tcaagccccg gctgctggaa gacctggagg agcagctgta ctgctcagcc
     3301 tttgaggagg cagccctgac ccgcaggatc tgtaatccta cttcctgctg gctgccttta
     3361 gacatggagc tgctacacag acaggtcctg gctctgcaga cacagagggt cctgctggag
     3421 aagaggcgga aggcttcagc ctggcaacgg aacctgggct accccctggc tatgctgtgc
     3481 ttgctggtgc tgacgggcct gtctgtgctc attgtggcca tccacatcct ggagctgctc
     3541 atcgatgagg ctgccatgcc ccgaggcatg cagggtacct ccttaggcca ggtctccttc
     3601 tccaagctgg gctcctttgg tgccgtcatt caggttgtac tcatctttta cctaatggtg
     3661 tcctcagttg tgggcttcta tagctctcca ctcttccgga gcctgcggcc cagatggcac
     3721 gacactgcca tgacgcagat aattgggaac tgtgtctgtc tcctggtcct aagctcagca
     3781 cttcctgtct tctctcgaac cctggggctc actcgctttg acctgctggg tgactttgga
     3841 cgcttcaact ggctgggcaa tttctacatt gtgttcctct acaacgcagc ctttgcaggc
     3901 ctcaccacac tctgtctggt gaagaccttc actgcagctg tgcgggcaga gctgatccgg
     3961 gcctttgggc tggacagact gccgctgccc gtctccggtt tcccccaggc atctaggaag
     4021 acccagcacc agtgactcga gtctagatgt acaaagtggt gataatcgaa ttccgataat
     4081 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     4141 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     4201 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     4261 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     4321 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     4381 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     4441 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     4501 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     4561 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     4621 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ggaattcccg
     4681 cggttcgaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct
     4741 ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc
     4801 acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct
     4861 actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga
     4921 caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg
     4981 aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct
     5041 cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg
     5101 ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc
     5161 gcgctgttct cctcttcctc atctccgggc ctttcgacct cacgtggcca ccatgaccga
     5221 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     5281 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     5341 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     5401 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     5461 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     5521 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     5581 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     5641 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     5701 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     5761 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaggtacctt
     5821 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     5881 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt
     5941 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     6001 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     6061 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     6121 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     6181 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6241 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6301 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct
     6361 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     6421 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     6481 aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc
     6541 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     6601 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     6661 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     6721 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     6781 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     6841 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     6901 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     6961 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     7021 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     7081 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     7141 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     7201 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     7261 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     7321 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     7381 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     7441 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     7501 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     7561 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     7621 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     7681 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     7741 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     7801 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     7861 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     7921 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     7981 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     8041 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     8101 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     8161 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     8221 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     8281 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     8341 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     8401 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     8461 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     8521 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     8581 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     8641 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     8701 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     8761 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     8821 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     8881 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     8941 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     9001 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     9061 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     9121 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     9181 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     9241 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     9301 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag
     9361 ctt
//
