LOCUS       Exported                9688 bp ds-DNA     circular SYN 07-AUG-2023
DEFINITION  Aguilera_Uribe_W6Y_1_T1     coverage: 1.22e+03x.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9688)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jun 12, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9688
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     polyA_signal    103..224
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             296..529
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     CDS             complement(657..1256)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     promoter        complement(1278..1788)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     misc_feature    1837..1954
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(1999..2022)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(2023..4353)
                     /label=TSC22D2 Full length
     promoter        complement(4391..4594)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             complement(5201..5245)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5430..5663
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    6156..6281
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6328..6508
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(6509..6735)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     promoter        complement(6763..6781)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6802..6818)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6826..6842
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(6850..6880)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6895..6916
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7204..7792)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7963..8823)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8824..8928)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8954..9409)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     9551..9567
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        9577..9595
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      join(9616..9688,1..63)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
       61 gatgggcgga gttaggggcg ggatagctag agccagacat gataagatac attgatgagt
      121 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
      181 ctattgcttt atttgtaacc attataagct gcaataaaca agttcctctc actctctgat
      241 attcatttct ttgcaagtta taaatactga ataataagat gacatgaact actactgcta
      301 gagattttcc acactgacta aaagggtctg agggatctct agttaccaga gtcacacaac
      361 agacgggcac acactacttg aagcactcaa ggcaagcttt attgaggctt aagcagtggg
      421 ttccctagtt agccagagag ctcccaggct cagatctggt ctaaccagag agacccagta
      481 caagcaaaaa gcagatcttg tcttcgttgg gagtgaatta gcccttccag tccccccttt
      541 tcttttaaaa agtggctaag atctacagct gccttgtaag tcattggtct taaaggtacc
      601 gatgcatggg gtcgtgcgct cctttcggtc gggcgctgcg ggtcgtgggg cgggcgtcag
      661 gcaccgggct tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg
      721 gtgacggtga agccgagccg ctcgtagaag gggaggttgc ggggcgcgga ggtctccagg
      781 aaggcgggca ccccggcgcg ctcggccgcc tccactccgg ggagcacgac ggcgctgccc
      841 agacccttgc cctggtggtc gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc
      901 ttgggccggt gcggcgccag gaggccttcc atctgttgct gcgcggccag ccgggaaccg
      961 ctcaactcgg ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc gacgctctcc
     1021 ggcgtggtcc agaccgccac cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg
     1081 agcccgacgc gcgtgaggaa gagttcttgc agctcggtga cccgctcgat gtggcggtcc
     1141 ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg
     1201 gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc ggtcatggta
     1261 agctccggtg gatccccctg gggagagagg tcggtgattc ggtcaacgag ggagccgact
     1321 gccgacgtgc gctccggagg cttgcagaat gcggaacacc gcgcgggcag gaacagggcc
     1381 cacactaccg ccccacaccc cgcctcccgc accgcccctt cccggccgct gctctcggcg
     1441 cgccctgctg agcagccgct attggccaca gcccatcgcg gtcggcgcgc tgccattgct
     1501 ccctggcgct gtccgtctgc gagggtacta gtgagacgtg cggcttccgt ttgtcacgtc
     1561 cggcacgccg cgaaccgcaa ggaaccttcc cgacttaggg gcggagcagg aagcgtcgcc
     1621 ggggggccca caagggtagc ggcgaagatc cgggtgacgc tgcgaacgga cgtgaagaat
     1681 gtgcgagacc cagggtcggc gccgctgcgt ttcccggaac cacgcccaga gcagccgcgt
     1741 ccctgcgcaa acccagggct gccttggaaa aggcgcaacc ccaaccccct cgagccgcgg
     1801 ccaaagtgga tctctgctgt ccctgtaata aacccgaaaa ttttgaattt ttgtaatttg
     1861 tttttgtaat tctttagttt gtatgtctgt tgctattatg tctactattc tttcccctgc
     1921 actgtacccc ccaatccccc cttttctttt aaaattgtgg atgaatactg ccatttgtct
     1981 cgaggtcgag aattcttact tgtcatcgtc gtccttgtag tctgctgagg agacattcgg
     2041 ctgctgtgga ggttgcgttg gctgcggagg ctgtgctatc actgcttgct gttgagaagt
     2101 gctaccagga ttggcctgtt gggttgggag ttgggataat tgatcattgc ttgaaagaga
     2161 ttttaacagt gcattttctc gttcaagtaa agagtttctt tcaactaatt cttttatttg
     2221 ttcctttaaa acttccactt cttctcttac tgcatacatc aaatggcttt tcaccagatc
     2281 cattgcttgt tctattttgt tgtcaatggc tacaacacct cccccagatg cactatccca
     2341 gaggctcttt gattccttta acgtgttcaa atggaacgct tggtagaaag cagttgaagg
     2401 attcctgtct tcatcaccat caacaggaat agctatgctg aatacagagg tggccagact
     2461 gttcataggt gttaactgaa gggggtttgc cagggaatct gcaacaggcg gcttcacaac
     2521 aggcttattt tcagcaatga gagaaagtgg tggttgaggt aggggttctg attttcttct
     2581 agtatcgtcg acttgcccga ctaaaggctg ggtctgagtt tgaaactggc ttaggtcaga
     2641 ctgtggtaga cttgttggtg cgctgtgtgt gcctggcgct aagggcagcc caacatgctg
     2701 gattatgctg ctgctcctgc tgactggcgt atggctgctc agctgttgcg actgggccag
     2761 aggcgcgggt acatttggca tagtaacaga agtggtagac acactaggca cgcttggagc
     2821 gggcacggct gcaggcacgt ttggaactcc gggcaccacg gcgtgagggc caccaggtac
     2881 ggctgacagc ggaccactgc cacccatctg cggcggaggc acggactggg gctgcccagc
     2941 cccggcagga ccgaggcacg gctgcaccac gcctcccaca gtagccgggg gcactccagt
     3001 cggctgacaa ggcgcgacct gcccgccttg cgctggtccc gtccgggggg ccatggcttc
     3061 gctgggctgg gaagacgcgc ccatgagctg cgcgcccacc gaggaagcat tctggccggt
     3121 gcccacggga agggtggccg ccgtggcggg gctcgctgcg gcgccggtgg acgagggctg
     3181 cgcggggctt ggcggctgca ggccggccac gtgctgctgc aggtactcac tctggccgga
     3241 gggctggacg ggcagcaaat gtccgggcgg tatctgaggc tgaggatacg cgaactgctg
     3301 gggctgctgc gccgcggggg cgcccactgc agggcccggc tgcggaggca gggacatagc
     3361 cggctgcgcc agggttacat tcgtcggcgg cagagtctgt cccccgggcc ccgctccctg
     3421 gggctggctg ggctgcgcgg ctgccatcat tggctgcggc ccggtcgcgg ctcccgggaa
     3481 cggcggcagc ggagccgagc tttgagccac agccccacct acgggtggcg gcggcggctg
     3541 tggctgccca acgctaaaac tctgcggctg ggccggagtg ggctggctca ttttctccga
     3601 cgggggtagc tgtgacacag cagtcaagga gctgtcagtt cccgactcgg gcccgtgcgc
     3661 cccctgcatg gaggccacta ctaccaccac cgaccctccg gtggcgccca ggccgctgtc
     3721 ccgctccgca gtctggtcaa aagtactgct gtgtctaatg caatccccgg atctagtcag
     3781 gacgctgctg tctgaatccc tctcatagta ttccatacac gtccatcggc cgcgtctata
     3841 gggctctccg ctcccgtggt ccagcttgat cacgcgaaaa cgggaactac atgtggtggg
     3901 aggctgagat ggggctgcag tcactggccc ggcggatgag cccgcgagct gggggccgcc
     3961 gggtgctccg ggggcggggg ccgaagtggc agccgccgcc agggtactgg cgagcgcccc
     4021 agacacgctc cgggccgaaa cgactcctcc tccgttggcg ggagcagcag ccgccgctgc
     4081 cagctgccca tctaggagga ggtttgggga gacggtcccg ggagtctccg catccccaac
     4141 attgttaagc gtctcttcgg aagagctgcg ctcgcagacc tcctcagggc cataatccgt
     4201 ggcccgagag acgtcgaaaa tctcggaaga gacgtcctct gtgcgtgact cgtccgggtc
     4261 gtccaagctc tcggtgtcct cggtgatgct agtggccacc tgggccgtgg tgacactggt
     4321 gatctggaag cagctcttct tcttggccgg catcttggac atgctagcgg atctgacggt
     4381 tcactaaacc agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg
     4441 tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca
     4501 aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc
     4561 cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt
     4621 agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg
     4681 gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatag tccattttaa
     4741 aacataattt taaaactgca aactacccaa gaaattatta ctttctacgt cacgtatttt
     4801 gtactaatat ctttgtgttt acagtcaaat taattccaat tatctctcta acagccttgt
     4861 atcgtatatg caaatatgaa ggaatcatgg gaaataggcc ctcggtgaag ggggcggccg
     4921 ctcgaggcta gtctcgtgat cgataccgtc gagatccgtt cactaatcga atggatctgt
     4981 ctctgtctct ctctccacct tcttcttcta ttccttcggg cctgtcgggt cccctcgggg
     5041 ttgggaggtg ggtctgaaac gataatggtg aatatccctg cctaactcta ttcactatag
     5101 aaagtacagc aaaaactatt cttaaaccta ccaagcctcc tactatcatt atgaataatt
     5161 ttatatacca cagccaattt gttatgttaa accaattcca caaacttgcc catttatcta
     5221 attccaataa ttcttgttca ttcttttctt gctggttttg cgattcttca attaaggagt
     5281 gtattaagct tgtgtaattg ttaatttctc tgtcccactc catccaggtc gtgtgattcc
     5341 aaatctgttc cagagattta ttactccaac tagcattcca aggcacagca gtggtgcaaa
     5401 tgagttttcc agagcaaccc caaatcccca ggagctgttg atcctttagg tatctttcca
     5461 cagccaggat tcttgcctgg agctgcttga tgccccagac tgtgagttgc aacagatgct
     5521 gttgcgcctc aatagccctc agcaaattgt tctgctgctg cactatacca gacaataatt
     5581 gtctggcctg taccgtcagc gtcattgacg ctgcgcccat agtgcttcct gctgctccca
     5641 agaacccaag gaacaaagct cctattccca ctgctctttt ttctctctgc accactcttc
     5701 tctttgcctt ggtgggtgct actcctaatg gttcaatttt tactacttta tatttatata
     5761 attcacttct ccaattgtcc ctcatatctc ctcctccagg tctgaagatc agcggccgct
     5821 tgctgtgcgg tggtcttact tttgttttgc tcttcctcta tcttgtctaa agcttccttg
     5881 gtgtctttta tctctatcct ttgatgcaca caatagaggg ttgctactgt attatataat
     5941 gatctaagtt cttctgatcc tgtctgaagg gatggttgta gctgtcccag tatttgtcta
     6001 cagccttctg atgtttctaa caggccagga ttaactgcga atcgttctag ctccctgctt
     6061 gcccatacta tatgttttaa tttatatttt ttctttcccc ctggccttaa ccgaattttt
     6121 tcccatcgcg atctaattct cccccgctta atactgacgc tctcgcaccc atctctctcc
     6181 ttctagcctc cgctagtcaa aatttttggc gtactcacca gtcgccgccc ctcgcctctt
     6241 gccgtgcgcg cttcagcaag ccgagtcctg cgtcgagaga gctcctctgg tttccctttc
     6301 gctttcaagt ccctgttcgg gcgccactgc tagagatttt ccacactgac taaaagggtc
     6361 tgagggatct ctagttacca gagtcacaca acagacgggc acacactact tgaagcactc
     6421 aaggcaagct ttattgaggc ttaagcagtg ggttccctag ttagccagag agctcccagg
     6481 ctcagatctg gtctaaccag agagacccgt ttatgtatcg agctaggcac ttaaatacaa
     6541 tatctctgca atgcggcaat tcagtggttc gtccaatcca tgtcagaccc gtctgttgcc
     6601 ttcctaataa ggcacgatcg taccacctta cttccaccaa tcggcatgca cggtgctttt
     6661 tctctccttg taaggcatgt tgctaactca tcgttaccat gttgcaagac tacaagagta
     6721 ttgcataaga ctacattaag cttgcagctc cagcttttgt tccctttagt gagggttaat
     6781 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6841 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6901 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6961 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     7021 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     7081 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     7141 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     7201 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     7261 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     7321 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     7381 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7441 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7501 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7561 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7621 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     7681 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7741 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7801 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7861 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7921 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7981 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     8041 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     8101 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     8161 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     8221 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     8281 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     8341 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     8401 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8461 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8521 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8581 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8641 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8701 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8761 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8821 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8881 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8941 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     9001 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     9061 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     9121 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     9181 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     9241 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     9301 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     9361 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
     9421 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     9481 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     9541 tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa
     9601 ttgggtacgt ccctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg
     9661 ccgaggcggc ctcggcctct gcataaat
//
