LOCUS       Exported                8944 bp ds-DNA     circular SYN 10-JUN-2024
DEFINITION  Aguilera_Uribe_W6Y_6_T6     coverage: 774x.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8944)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jun 12, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8944
                     /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"
     misc_feature    complement(2023..3618)
                     /label=TSC22D2 C-term
     promoter        complement(3647..3850)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             complement(4457..4501)
                     /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    4686..4919
                     /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    5412..5537
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5584..5764
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(5765..5991)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     promoter        complement(6019..6037)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6058..6074)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6082..6098
                     /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(6106..6136)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6151..6172
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6460..7048)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7219..8079)
                     /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(8080..8184)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8210..8665)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     8807..8823
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8833..8851
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      join(8872..8944,1..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 tgtgacatgc tagcggatct gacggttcac taaaccagct ctgcttatat
     3661 agacctccca ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac
     3721 attttggaaa gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt
     3781 ggagacttgg aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa
     3841 accgcatcac catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt
     3901 cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa
     3961 tagggggcgt acttggcata tgatagtcca ttttaaaaca taattttaaa actgcaaact
     4021 acccaagaaa ttattacttt ctacgtcacg tattttgtac taatatcttt gtgtttacag
     4081 tcaaattaat tccaattatc tctctaacag ccttgtatcg tatatgcaaa tatgaaggaa
     4141 tcatgggaaa taggccctcg gtgaaggggg cggccgctcg aggctagtct cgtgatcgat
     4201 accgtcgaga tccgttcact aatcgaatgg atctgtctct gtctctctct ccaccttctt
     4261 cttctattcc ttcgggcctg tcgggtcccc tcggggttgg gaggtgggtc tgaaacgata
     4321 atggtgaata tccctgccta actctattca ctatagaaag tacagcaaaa actattctta
     4381 aacctaccaa gcctcctact atcattatga ataattttat ataccacagc caatttgtta
     4441 tgttaaacca attccacaaa cttgcccatt tatctaattc caataattct tgttcattct
     4501 tttcttgctg gttttgcgat tcttcaatta aggagtgtat taagcttgtg taattgttaa
     4561 tttctctgtc ccactccatc caggtcgtgt gattccaaat ctgttccaga gatttattac
     4621 tccaactagc attccaaggc acagcagtgg tgcaaatgag ttttccagag caaccccaaa
     4681 tccccaggag ctgttgatcc tttaggtatc tttccacagc caggattctt gcctggagct
     4741 gcttgatgcc ccagactgtg agttgcaaca gatgctgttg cgcctcaata gccctcagca
     4801 aattgttctg ctgctgcact ataccagaca ataattgtct ggcctgtacc gtcagcgtca
     4861 ttgacgctgc gcccatagtg cttcctgctg ctcccaagaa cccaaggaac aaagctccta
     4921 ttcccactgc tcttttttct ctctgcacca ctcttctctt tgccttggtg ggtgctactc
     4981 ctaatggttc aatttttact actttatatt tatataattc acttctccaa ttgtccctca
     5041 tatctcctcc tccaggtctg aagatcagcg gccgcttgct gtgcggtggt cttacttttg
     5101 ttttgctctt cctctatctt gtctaaagct tccttggtgt cttttatctc tatcctttga
     5161 tgcacacaat agagggttgc tactgtatta tataatgatc taagttcttc tgatcctgtc
     5221 tgaagggatg gttgtagctg tcccagtatt tgtctacagc cttctgatgt ttctaacagg
     5281 ccaggattaa ctgcgaatcg ttctagctcc ctgcttgccc atactatatg ttttaattta
     5341 tattttttct ttccccctgg ccttaaccga attttttccc atcgcgatct aattctcccc
     5401 cgcttaatac tgacgctctc gcacccatct ctctccttct agcctccgct agtcaaaatt
     5461 tttggcgtac tcaccagtcg ccgcccctcg cctcttgccg tgcgcgcttc agcaagccga
     5521 gtcctgcgtc gagagagctc ctctggtttc cctttcgctt tcaagtccct gttcgggcgc
     5581 cactgctaga gattttccac actgactaaa agggtctgag ggatctctag ttaccagagt
     5641 cacacaacag acgggcacac actacttgaa gcactcaagg caagctttat tgaggcttaa
     5701 gcagtgggtt ccctagttag ccagagagct cccaggctca gatctggtct aaccagagag
     5761 acccgtttat gtatcgagct aggcacttaa atacaatatc tctgcaatgc ggcaattcag
     5821 tggttcgtcc aatccatgtc agacccgtct gttgccttcc taataaggca cgatcgtacc
     5881 accttacttc caccaatcgg catgcacggt gctttttctc tccttgtaag gcatgttgct
     5941 aactcatcgt taccatgttg caagactaca agagtattgc ataagactac attaagcttg
     6001 cagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     6061 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6121 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6181 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6241 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6301 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6361 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6421 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6481 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6541 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6601 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6661 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6721 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6781 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6841 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6901 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6961 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7021 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7081 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7141 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7201 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7261 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7321 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7381 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7441 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7501 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7561 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7621 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7681 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7741 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7801 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7861 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7921 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7981 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8041 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8101 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8161 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
     8221 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     8281 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     8341 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     8401 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     8461 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     8521 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     8581 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     8641 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa
     8701 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     8761 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     8821 agtgagcgcg cgtaatacga ctcactatag ggcgaattgg gtacgtccct aggcctccaa
     8881 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
     8941 aaat
//
