LOCUS       Exported                8945 bp ds-DNA     circular SYN 10-JUN-2024
DEFINITION  Aguilera_Uribe_W6Y_5_T5     coverage: 693x.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8945)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jun 12, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8945
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     polyA_signal    103..224
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             296..530
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     CDS             complement(658..1257)
                     /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(1279..1789)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     misc_feature    1838..1955
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    complement(1997..3592)
                     /label=TSC22D2 C-term
     CDS             complement(3593..3616)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3648..3851)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             complement(4458..4502)
                     /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    4687..4920
                     /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    5413..5538
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5585..5765
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(5766..5992)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     promoter        complement(6020..6038)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6059..6075)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6083..6099
                     /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(6107..6137)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6152..6173
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6461..7049)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7220..8080)
                     /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(8081..8185)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8211..8666)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     8808..8824
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8834..8852
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      join(8873..8945,1..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 agggattctc tagttaccag agtcacacaa
      361 cagacgggca cacactactt gaagcactca aggcaagctt tattgaggct taagcagtgg
      421 gttccctagt tagccagaga gctcccaggc tcagatctgg tctaaccaga gagacccagt
      481 acaagcaaaa agcagatctt gtcttcgttg ggagtgaatt agcccttcca gtcccccctt
      541 ttcttttaaa aagtggctaa gatctacagc tgccttgtaa gtcattggtc ttaaaggtac
      601 cgatgcatgg ggtcgtgcgc tcctttcggt cgggcgctgc gggtcgtggg gcgggcgtca
      661 ggcaccgggc ttgcgggtca tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc
      721 ggtgacggtg aagccgagcc gctcgtagaa ggggaggttg cggggcgcgg aggtctccag
      781 gaaggcgggc accccggcgc gctcggccgc ctccactccg gggagcacga cggcgctgcc
      841 cagacccttg ccctggtggt cgggcgagac gccgacggtg gccaggaacc acgcgggctc
      901 cttgggccgg tgcggcgcca ggaggccttc catctgttgc tgcgcggcca gccgggaacc
      961 gctcaactcg gccatgcgcg ggccgatctc ggcgaacacc gcccccgctt cgacgctctc
     1021 cggcgtggtc cagaccgcca ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc
     1081 gagcccgacg cgcgtgagga agagttcttg cagctcggtg acccgctcga tgtggcggtc
     1141 cggatcgacg gtgtggcgcg tggcggggta gtcggcgaac gcggcggcga gggtgcgtac
     1201 ggccctgggg acgtcgtcgc gggtggcgag gcgcaccgtg ggcttgtact cggtcatggt
     1261 aagctccggt ggatccccct ggggagagag gtcggtgatt cggtcaacga gggagccgac
     1321 tgccgacgtg cgctccggag gcttgcagaa tgcggaacac cgcgcgggca ggaacagggc
     1381 ccacactacc gccccacacc ccgcctcccg caccgcccct tcccggccgc tgctctcggc
     1441 gcgccctgct gagcagccgc tattggccac agcccatcgc ggtcggcgcg ctgccattgc
     1501 tccctggcgc tgtccgtctg cgagggtact agtgagacgt gcggcttccg tttgtcacgt
     1561 ccggcacgcc gcgaaccgca aggaaccttc ccgacttagg ggcggagcag gaagcgtcgc
     1621 cggggggccc acaagggtag cggcgaagat ccgggtgacg ctgcgaacgg acgtgaagaa
     1681 tgtgcgagac ccagggtcgg cgccgctgcg tttcccggaa ccacgcccag agcagccgcg
     1741 tccctgcgca aacccagggc tgccttggaa aaggcgcaac cccaaccccc tcgagccgcg
     1801 gccaaagtgg atctctgctg tccctgtaat aaacccgaaa attttgaatt tttgtaattt
     1861 gtttttgtaa ttctttagtt tgtatgtctg ttgctattat gtctactatt ctttcccctg
     1921 cactgtaccc cccaatcccc ccttttcttt taaaattgtg gatgaatact gccatttgtc
     1981 tcgaggtcga gaattcttat gctgaggaga cattcggctg ctgtggaggt tgcgttggct
     2041 gcggaggctg tgctatcact gcttgctgtt gagaagtgct accaggattg gcctgttggg
     2101 ttgggagttg ggataattga tcattgcttg aaagagattt taacagtgca ttttctcgtt
     2161 caagtaaaga gtttctttca actaattctt ttatttgttc ctttaaaact tccacttctt
     2221 ctcttactgc atacatcaaa tggcttttca ccagatccat tgcttgttct attttgttgt
     2281 caatggctac aacacctccc ccagatgcac tatcccagag gctctttgat tcctttaacg
     2341 tgttcaaatg gaacgcttgg tagaaagcag ttgaaggatt cctgtcttca tcaccatcaa
     2401 caggaatagc tatgctgaat acagaggtgg ccagactgtt cataggtgtt aactgaaggg
     2461 ggtttgccag ggaatctgca acaggcggct tcacaacagg cttattttca gcaatgagag
     2521 aaagtggtgg ttgaggtagg ggttctgatt ttcttctagt atcgtcgact tgcccgacta
     2581 aaggctgggt ctgagtttga aactggctta ggtcagactg tggtagactt gttggtgcgc
     2641 tgtgtgtgcc tggcgctaag ggcagcccaa catgctggat tatgctgctg ctcctgctga
     2701 ctggcgtatg gctgctcagc tgttgcgact gggccagagg cgcgggtaca tttggcatag
     2761 taacagaagt ggtagacaca ctaggcacgc ttggagcggg cacggctgca ggcacgtttg
     2821 gaactccggg caccacggcg tgagggccac caggtacggc tgacagcgga ccactgccac
     2881 ccatctgcgg cggaggcacg gactggggct gcccagcccc ggcaggaccg aggcacggct
     2941 gcaccacgcc tcccacagta gccgggggca ctccagtcgg ctgacaaggc gcgacctgcc
     3001 cgccttgcgc tggtcccgtc cggggggcca tggcttcgct gggctgggaa gacgcgccca
     3061 tgagctgcgc gcccaccgag gaagcattct ggccggtgcc cacgggaagg gtggccgccg
     3121 tggcggggct cgctgcggcg ccggtggacg agggctgcgc ggggcttggc ggctgcaggc
     3181 cggccacgtg ctgctgcagg tactcactct ggccggaggg ctggacgggc agcaaatgtc
     3241 cgggcggtat ctgaggctga ggatacgcga actgctgggg ctgctgcgcc gcgggggcgc
     3301 ccactgcagg gcccggctgc ggaggcaggg acatagccgg ctgcgccagg gttacattcg
     3361 tcggcggcag agtctgtccc ccgggccccg ctccctgggg ctggctgggc tgcgcggctg
     3421 ccatcattgg ctgcggcccg gtcgcggctc ccgggaacgg cggcagcgga gccgagcttt
     3481 gagccacagc cccacctacg ggtggcggcg gcggctgtgg ctgcccaacg ctaaaactct
     3541 gcggctgggc cggagtgggc tggctcattt tctccgacgg gggtagctgt gacttgtcat
     3601 cgtcgtcctt gtagtccatg ctagcggatc tgacggttca ctaaaccagc tctgcttata
     3661 tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga
     3721 cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg
     3781 tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa
     3841 aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag
     3901 tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca
     3961 atagggggcg tacttggcat atgatagtcc attttaaaac ataattttaa aactgcaaac
     4021 tacccaagaa attattactt tctacgtcac gtattttgta ctaatatctt tgtgtttaca
     4081 gtcaaattaa ttccaattat ctctctaaca gccttgtatc gtatatgcaa atatgaagga
     4141 atcatgggaa ataggccctc ggtgaagggg gcggccgctc gaggctagtc tcgtgatcga
     4201 taccgtcgag atccgttcac taatcgaatg gatctgtctc tgtctctctc tccaccttct
     4261 tcttctattc cttcgggcct gtcgggtccc ctcggggttg ggaggtgggt ctgaaacgat
     4321 aatggtgaat atccctgcct aactctattc actatagaaa gtacagcaaa aactattctt
     4381 aaacctacca agcctcctac tatcattatg aataatttta tataccacag ccaatttgtt
     4441 atgttaaacc aattccacaa acttgcccat ttatctaatt ccaataattc ttgttcattc
     4501 ttttcttgct ggttttgcga ttcttcaatt aaggagtgta ttaagcttgt gtaattgtta
     4561 atttctctgt cccactccat ccaggtcgtg tgattccaaa tctgttccag agatttatta
     4621 ctccaactag cattccaagg cacagcagtg gtgcaaatga gttttccaga gcaaccccaa
     4681 atccccagga gctgttgatc ctttaggtat ctttccacag ccaggattct tgcctggagc
     4741 tgcttgatgc cccagactgt gagttgcaac agatgctgtt gcgcctcaat agccctcagc
     4801 aaattgttct gctgctgcac tataccagac aataattgtc tggcctgtac cgtcagcgtc
     4861 attgacgctg cgcccatagt gcttcctgct gctcccaaga acccaaggaa caaagctcct
     4921 attcccactg ctcttttttc tctctgcacc actcttctct ttgccttggt gggtgctact
     4981 cctaatggtt caatttttac tactttatat ttatataatt cacttctcca attgtccctc
     5041 atatctcctc ctccaggtct gaagatcagc ggccgcttgc tgtgcggtgg tcttactttt
     5101 gttttgctct tcctctatct tgtctaaagc ttccttggtg tcttttatct ctatcctttg
     5161 atgcacacaa tagagggttg ctactgtatt atataatgat ctaagttctt ctgatcctgt
     5221 ctgaagggat ggttgtagct gtcccagtat ttgtctacag ccttctgatg tttctaacag
     5281 gccaggatta actgcgaatc gttctagctc cctgcttgcc catactatat gttttaattt
     5341 atattttttc tttccccctg gccttaaccg aattttttcc catcgcgatc taattctccc
     5401 ccgcttaata ctgacgctct cgcacccatc tctctccttc tagcctccgc tagtcaaaat
     5461 ttttggcgta ctcaccagtc gccgcccctc gcctcttgcc gtgcgcgctt cagcaagccg
     5521 agtcctgcgt cgagagagct cctctggttt ccctttcgct ttcaagtccc tgttcgggcg
     5581 ccactgctag agattttcca cactgactaa aagggtctga gggatctcta gttaccagag
     5641 tcacacaaca gacgggcaca cactacttga agcactcaag gcaagcttta ttgaggctta
     5701 agcagtgggt tccctagtta gccagagagc tcccaggctc agatctggtc taaccagaga
     5761 gacccgttta tgtatcgagc taggcactta aatacaatat ctctgcaatg cggcaattca
     5821 gtggttcgtc caatccatgt cagacccgtc tgttgccttc ctaataaggc acgatcgtac
     5881 caccttactt ccaccaatcg gcatgcacgg tgctttttct ctccttgtaa ggcatgttgc
     5941 taactcatcg ttaccatgtt gcaagactac aagagtattg cataagacta cattaagctt
     6001 gcagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     6061 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6121 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6181 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6241 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6301 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6361 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6421 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6481 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6541 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6601 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6661 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6721 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6781 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6841 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6901 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6961 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7021 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7081 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7141 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7201 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7261 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7321 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7381 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7441 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7501 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7561 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7621 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7681 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7741 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7801 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7861 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7921 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7981 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8041 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8101 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8161 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag
     8221 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     8281 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     8341 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     8401 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     8461 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     8521 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     8581 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     8641 gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga
     8701 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     8761 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     8821 cagtgagcgc gcgtaatacg actcactata gggcgaattg ggtacgtccc taggcctcca
     8881 aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca
     8941 taaat
//
