LOCUS       V022208                 9425 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022208
VERSION     V022208
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..4350
                     /label="TGFBI(NM_000358)"
                     /note="TGFBI(NM_000358)"
                     /gene="TGFBI"
     promoter        4395..4606
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4619..4887
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4914..5513
                     /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    5520..6108
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6182..6415
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6487..6608
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6627..6762
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6800..6816)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6824..6840
                     /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(6848..6878)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6893..6914
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7202..7790)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7961..8821)
                     /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(8822..8926)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9400..9416
                     /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 catggcgctc ttcgtgcggc tgctggctct cgccctggct
     2341 ctggccctgg gccccgccgc gaccctggcg ggtcccgcca agtcgcccta ccagctggtg
     2401 ctgcagcaca gcaggctccg gggccgccag cacggcccca acgtgtgtgc tgtgcagaag
     2461 gttattggca ctaataggaa gtacttcacc aactgcaagc agtggtacca aaggaaaatc
     2521 tgtggcaaat caacagtcat cagctacgag tgctgtcctg gatatgaaaa ggtccctggg
     2581 gagaagggct gtccagcagc cctaccactc tcaaaccttt acgagaccct gggagtcgtt
     2641 ggatccacca ccactcagct gtacacggac cgcacggaga agctgaggcc tgagatggag
     2701 gggcccggca gcttcaccat cttcgcccct agcaacgagg cctgggcctc cttgccagct
     2761 gaagtgctgg actccctggt cagcaatgtc aacattgagc tgctcaatgc cctccgctac
     2821 catatggtgg gcaggcgagt cctgactgat gagctgaaac acggcatgac cctcacctct
     2881 atgtaccaga attccaacat ccagatccac cactatccta atgggattgt aactgtgaac
     2941 tgtgcccggc tgctgaaagc cgaccaccat gcaaccaacg gggtggtgca cctcatcgat
     3001 aaggtcatct ccaccatcac caacaacatc cagcagatca ttgagatcga ggacaccttt
     3061 gagacccttc gggctgctgt ggctgcatca gggctcaaca cgatgcttga aggtaacggc
     3121 cagtacacgc ttttggcccc gaccaatgag gccttcgaga agatccctag tgagactttg
     3181 aaccgtatcc tgggcgaccc agaagccctg agagacctgc tgaacaacca catcttgaag
     3241 tcagctatgt gtgctgaagc catcgttgcg gggctgtctg tagagaccct ggagggcacg
     3301 acactggagg tgggctgcag cggggacatg ctcactatca acgggaaggc gatcatctcc
     3361 aataaagaca tcctagccac caacggggtg atccactaca ttgatgagct actcatccca
     3421 gactcagcca agacactatt tgaattggct gcagagtctg atgtgtccac agccattgac
     3481 cttttcagac aagccggcct cggcaatcat ctctctggaa gtgagcggtt gaccctcctg
     3541 gctcccctga attctgtatt caaagatgga acccctccaa ttgatgccca tacaaggaat
     3601 ttgcttcgga accacataat taaagaccag ctggcctcta agtatctgta ccatggacag
     3661 accctggaaa ctctgggcgg caaaaaactg agagtttttg tttatcgtaa tagcctctgc
     3721 attgagaaca gctgcatcgc ggcccacgac aagaggggga ggtacgggac cctgttcacg
     3781 atggaccggg tgctgacccc cccaatgggg actgtcatgg atgtcctgaa gggagacaat
     3841 cgctttagca tgctggtagc tgccatccag tctgcaggac tgacggagac cctcaaccgg
     3901 gaaggagtct acacagtctt tgctcccaca aatgaagcct tccgagccct gccaccaaga
     3961 gaacggagca gactcttggg agatgccaag gaacttgcca acatcctgaa ataccacatt
     4021 ggtgatgaaa tcctggttag cggaggcatc ggggccctgg tgcggctaaa gtctctccaa
     4081 ggtgacaagc tggaagtcag cttgaaaaac aatgtggtga gtgtcaacaa ggagcctgtt
     4141 gccgagcctg acatcatggc cacaaatggc gtggtccatg tcatcaccaa tgttctgcag
     4201 cctccagcca acagacctca ggaaagaggg gatgaacttg cagactctgc gcttgagatc
     4261 ttcaaacaag catcagcgtt ttccagggct tcccagaggt ctgtgcgact agcccctgtc
     4321 tatcaaaagt tattagagag gatgaagcat taggcggccg cgaaggatct gcgatcgctc
     4381 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     4441 ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     4501 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     4561 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg
     4621 ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt
     4681 gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt
     4741 aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag
     4801 actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc
     4861 gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagatgaccg
     4921 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc
     4981 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca
     5041 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     5101 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     5161 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     5221 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     5281 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     5341 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     5401 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     5461 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca
     5521 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     5581 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     5641 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     5701 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     5761 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     5821 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     5881 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     5941 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     6001 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     6061 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta
     6121 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     6181 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct
     6241 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     6301 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     6361 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     6421 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     6481 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     6541 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     6601 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt
     6661 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     6721 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg
     6781 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6841 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6901 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6961 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     7021 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     7081 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     7141 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     7201 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     7261 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7321 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7381 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7441 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7501 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7561 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7621 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     7681 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7741 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7801 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7861 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7921 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7981 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     8041 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     8101 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     8161 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     8221 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     8281 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     8341 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8401 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8461 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8521 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8581 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8641 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8701 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8761 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8821 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8881 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8941 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     9001 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     9061 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     9121 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     9181 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     9241 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     9301 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     9361 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca
     9421 agctg
//
