LOCUS       V020749                 9955 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020749
VERSION     V020749
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             2295..4874
                     /label="LDLR(NM_000527)"
                     /note="LDLR(NM_000527)"
                     /gene="LDLR"
     promoter        4925..5136
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5149..5417
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5444..6043
                     /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    6050..6638
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6712..6945
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7017..7138
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7157..7292
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7330..7346)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7354..7370
                     /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(7378..7408)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7423..7444
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7732..8320)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8491..9351)
                     /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(9352..9456)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9930..9946
                     /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 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta
     2281 gagctagcgc caccatgggg ccctggggct ggaaattgcg ctggaccgtc gccttgctcc
     2341 tcgccgcggc ggggactgca gtgggcgaca gatgcgaaag aaacgagttc cagtgccaag
     2401 acgggaaatg catctcctac aagtgggtct gcgatggcag cgctgagtgc caggatggct
     2461 ctgatgagtc ccaggagacg tgcttgtctg tcacctgcaa atccggggac ttcagctgtg
     2521 ggggccgtgt caaccgctgc attcctcagt tctggaggtg cgatggccaa gtggactgcg
     2581 acaacggctc agacgagcaa ggctgtcccc ccaagacgtg ctcccaggac gagtttcgct
     2641 gccacgatgg gaagtgcatc tctcggcagt tcgtctgtga ctcagaccgg gactgcttgg
     2701 acggctcaga cgaggcctcc tgcccggtgc tcacctgtgg tcccgccagc ttccagtgca
     2761 acagctccac ctgcatcccc cagctgtggg cctgcgacaa cgaccccgac tgcgaagatg
     2821 gctcggatga gtggccgcag cgctgtaggg gtctttacgt gttccaaggg gacagtagcc
     2881 cctgctcggc cttcgagttc cactgcctaa gtggcgagtg catccactcc agctggcgct
     2941 gtgatggtgg ccccgactgc aaggacaaat ctgacgagga aaactgcgct gtggccacct
     3001 gtcgccctga cgaattccag tgctctgatg gaaactgcat ccatggcagc cggcagtgtg
     3061 accgggaata tgactgcaag gacatgagcg atgaagttgg ctgcgttaat gtgacactct
     3121 gcgagggacc caacaagttc aagtgtcaca gcggcgaatg catcaccctg gacaaagtct
     3181 gcaacatggc tagagactgc cgggactggt cagatgaacc catcaaagag tgcgggacca
     3241 acgaatgctt ggacaacaac ggcggctgtt cccacgtctg caatgacctt aagatcggct
     3301 acgagtgcct gtgccccgac ggcttccagc tggtggccca gcgaagatgc gaagatatcg
     3361 atgagtgtca ggatcccgac acctgcagcc agctctgcgt gaacctggag ggtggctaca
     3421 agtgccagtg tgaggaaggc ttccagctgg acccccacac gaaggcctgc aaggctgtgg
     3481 gctccatcgc ctacctcttc ttcaccaacc ggcacgaggt caggaagatg acgctggacc
     3541 ggagcgagta caccagcctc atccccaacc tgaggaacgt ggtcgctctg gacacggagg
     3601 tggccagcaa tagaatctac tggtctgacc tgtcccagag aatgatctgc agcacccagc
     3661 ttgacagagc ccacggcgtc tcttcctatg acaccgtcat cagcagagac atccaggccc
     3721 ccgacgggct ggctgtggac tggatccaca gcaacatcta ctggaccgac tctgtcctgg
     3781 gcactgtctc tgttgcggat accaagggcg tgaagaggaa aacgttattc agggagaacg
     3841 gctccaagcc aagggccatc gtggtggatc ctgttcatgg cttcatgtac tggactgact
     3901 ggggaactcc cgccaagatc aagaaagggg gcctgaatgg tgtggacatc tactcgctgg
     3961 tgactgaaaa cattcagtgg cccaatggca tcaccctaga tctcctcagt ggccgcctct
     4021 actgggttga ctccaaactt cactccatct caagcatcga tgtcaacggg ggcaaccgga
     4081 agaccatctt ggaggatgaa aagaggctgg cccacccctt ctccttggcc gtctttgagg
     4141 acaaagtatt ttggacagat atcatcaacg aagccatttt cagtgccaac cgcctcacag
     4201 gttccgatgt caacttgttg gctgaaaacc tactgtcccc agaggatatg gttctcttcc
     4261 acaacctcac ccagccaaga ggagtgaact ggtgtgagag gaccaccctg agcaatggcg
     4321 gctgccagta tctgtgcctc cctgccccgc agatcaaccc ccactcgccc aagtttacct
     4381 gcgcctgccc ggacggcatg ctgctggcca gggacatgag gagctgcctc acagaggctg
     4441 aggctgcagt ggccacccag gagacatcca ccgtcaggct aaaggtcagc tccacagccg
     4501 taaggacaca gcacacaacc acccgacctg ttcccgacac ctcccggctg cctggggcca
     4561 cccctgggct caccacggtg gagatagtga caatgtctca ccaagctctg ggcgacgttg
     4621 ctggcagagg aaatgagaag aagcccagta gcgtgagggc tctgtccatt gtcctcccca
     4681 tcgtgctcct cgtcttcctt tgcctggggg tcttccttct atggaagaac tggcggctta
     4741 agaacatcaa cagcatcaac tttgacaacc ccgtctatca gaagaccaca gaggatgagg
     4801 tccacatttg ccacaaccag gacggctaca gctacccctc gagacagatg gtcagtctgg
     4861 aggatgacgt ggcgtgagga tccgcggccg cgaaggatct gcgatcgctc cggtgcccgt
     4921 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     4981 tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     5041 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     5101 gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc
     5161 tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt
     5221 ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag
     5281 ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg
     5341 ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt
     5401 ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagatgaccg agtacaagcc
     5461 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc
     5521 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt
     5581 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     5641 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     5701 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     5761 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     5821 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     5881 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     5941 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     6001 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca atcaacctct
     6061 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     6121 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     6181 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     6241 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     6301 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     6361 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     6421 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     6481 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     6541 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     6601 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta agaccaatga
     6661 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     6721 taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct ctctggttag
     6781 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     6841 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     6901 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     6961 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact
     7021 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     7081 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     7141 atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     7201 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     7261 ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg gcccaaattc
     7321 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     7381 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     7441 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     7501 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     7561 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     7621 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     7681 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     7741 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7801 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7861 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7921 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7981 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     8041 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     8101 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     8161 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     8221 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     8281 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     8341 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     8401 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     8461 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     8521 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     8581 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     8641 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8701 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8761 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8821 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8881 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8941 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     9001 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     9061 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     9121 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     9181 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     9241 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     9301 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     9361 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     9421 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     9481 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     9541 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     9601 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     9661 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     9721 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     9781 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     9841 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     9901 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctg
//
