LOCUS       V029107                 8835 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029107
VERSION     V029107
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"
     regulatory      2288..2297
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2297..2362
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2369..3760
                     /label="ALG1(NM_019109)"
                     /note="ALG1(NM_019109)"
                     /gene="ALG1"
     promoter        3805..4016
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4029..4297
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4324..4923
                     /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    4930..5518
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5592..5825
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5897..6018
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6037..6172
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6210..6226)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6234..6250
                     /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(6258..6288)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6303..6324
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6612..7200)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7371..8231)
                     /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(8232..8336)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8810..8826
                     /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 agagctagcc accatggact acaaagacca tgacggtgat tataaagatc atgacatcga
     2341 ctacaaggat gacgatgaca aggaattcat ggcggcctca tgcttggtcc tgctggcgct
     2401 gtgtctgctg ctgccgctgc tgctgctggg aggatggaag cgctggcgcc gggggcgggc
     2461 ggcccggcat gtagtagcgg tggtgctggg cgacgtgggc cgcagccccc gtatgcagta
     2521 ccacgcgctg tcgttggcca tgcacggctt ctcggtgacc ctcctggggt tctgcaactc
     2581 caaaccccat gatgagctct tgcagaacaa cagaattcag attgtggggt tgacagaact
     2641 tcagagtctt gcagttgggc cccgagtttt ccagtacgga gtcaaagttg tacttcaggc
     2701 tatgtacttg ctgtggaagt tgatgtggag ggagccaggt gcctatatct ttctccagaa
     2761 ccccccaggt ctgcctagca ttgctgtctg ctggttcgtg ggctgccttt gtggaagcaa
     2821 gctcgtcatt gactggcaca actatggcta ctccatcatg ggtctggtgc atggccccaa
     2881 ccatcccctc gttctgctgg ccaagtggta cgagaagttc tttgggcgcc tgtcccacct
     2941 gaacctgtgt gttaccaatg ctatgcgaga agacctggcg gataactggc acatcagggc
     3001 tgtgaccgtc tacgacaagc ccgcatcttt ctttaaagag acacctctgg acctgcagca
     3061 ccggctcttc atgaagctgg gcagcatgca ctctccgttc agggcccgct cagaacctga
     3121 ggacccagtc acggagcggt cggccttcac ggagcgggat gctgggagcg ggctggtgac
     3181 gcgtctccgt gagcggccag ccctgctggt cagcagcacg agctggacag aggacgaaga
     3241 cttctccatc ctgctggcag ctttagaaaa gtttgaacaa ctgactcttg atggacacaa
     3301 ccttccttct ctcgtctgtg tgataacagg caaagggcct ctgagggagt attatagccg
     3361 cctcatccac cagaagcact tccagcacat ccaggtctgc accccctggc tggaggccga
     3421 ggactacccc ctgcttctag ggtcggcgga cctgggtgtc tgtctgcaca cgtcctccag
     3481 tggcctggac ctgcccatga aggtggtgga catgttcggg tgctgtttgc ctgtgtgtgc
     3541 tgtgaacttc aagtgtttac atgagctggt gaaacatgaa gaaaatggcc tggtctttga
     3601 ggactcagag gaactggcag ctcagctgca gatgcttttc tcaaactttc ctgatcctgc
     3661 gggcaagcta aaccagttcc ggaagaacct gcgggagtcg cagcagctcc gatgggatga
     3721 gagctgggtg cagactgtgc tccctttggt tatggacaca taagcggccg cgaaggatct
     3781 gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
     3841 tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg
     3901 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
     3961 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga
     4021 agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc
     4081 cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg
     4141 ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga
     4201 gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg
     4261 tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc
     4321 tagatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc
     4381 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg
     4441 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     4501 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     4561 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     4621 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     4681 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     4741 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     4801 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     4861 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     4921 tgagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     4981 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     5041 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     5101 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     5161 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     5221 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     5281 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     5341 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     5401 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     5461 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     5521 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     5581 aaagggggga ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg
     5641 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     5701 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     5761 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     5821 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     5881 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5941 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6001 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt
     6061 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     6121 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt
     6181 tgcagagacg gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     6241 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     6301 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6361 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6421 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6481 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6541 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6601 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6661 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6721 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6781 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6841 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6901 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6961 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7021 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     7081 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7141 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7201 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7261 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7321 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7381 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7441 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7501 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7561 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7621 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7681 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7741 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7801 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7861 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7921 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7981 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8041 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8101 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8161 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8221 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8281 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8341 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     8401 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     8461 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     8521 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     8581 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     8641 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
     8701 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     8761 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     8821 gccagtgcca agctg
//
