LOCUS       Exported                8614 bp ds-DNA     circular SYN 10-JUN-2024
DEFINITION  Aguilera_Uribe_W6Y_8_N2     coverage: 1.03e+03x.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8614)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jun 12, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8614
                     /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..3262)
                     /codon_start=2
                     /label=NRBP1 N-term
                     /translation="SEGESQTVLSSGSDPKVESSSSAPGLTSVSPPVTSTTSAASPEEE
                     EESEDESEILEESPCGRWQKRREEVNQRNVPGIDSAYLAMDTEEGVEVVWNEVQFSERK
                     NYKLQEEKVRAVFDNLIQLEHLNIVKFHKYWADIKENKARVIFITEYMSSGSLKQFLKK
                     TKKNHKTMNEKAWKRWCTQILSALSYLHSCDPPIIHGNLTCDTIFIQHNGLIKIGSVAP
                     DTINNHVKTCREEQKNLHFFAPEYGEVTNVTTAVDIYSFGMCALEMAVLEIQGNGESSY
                     VPQEAISSAIQLLEDPLQREFIQKCLQSEPARRPTARELLFHPALFEVPSLKLLAAHCI
                     VGHQHMIPENALEEITKNMDTSAVLAEIPAGPGREPVQTLYSQSPALELDKFLEDVRNG
                     IYPLTAFGLPRPQQPQQEEVTS"
     CDS             complement(3262..3285)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3317..3520)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             complement(4127..4171)
                     /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    4356..4589
                     /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    5082..5207
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5254..5434
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(5435..5661)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     promoter        complement(5689..5707)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    5715..5734
                     /label=N terminal HA- R 3' Gibson
     misc_feature    5715..5734
                     /label=C terminal HA- R 3' Gibson
     primer_bind     complement(5728..5744)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5752..5768
                     /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(5776..5806)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5821..5842
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6130..6718)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6889..7749)
                     /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(7750..7854)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7880..8335)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     misc_feature    8475..8494
                     /label=N terminal HA- L 5' Gibson
     misc_feature    8475..8494
                     /label=C terminal HA-L 5' Gibson
     primer_bind     8477..8493
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8503..8521
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      join(8542..8614,1..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 aattcctatg atgtcacctc ctcctgctgt ggctgctggg gccgaggcag
     2041 cccaaaggct gtcagaggat agatcccatt cctgacatct tcaaggaatt tatctaattc
     2101 cagagctggt gactgagagt acaaagtctg aactggttct cttcctggtc ctgcagggat
     2161 ttcagccagt acggcactag tatccatgtt tttggtgatc tcctctagag cgttctctgg
     2221 gatcatgtgt tggtgtccca caatgcagtg ggccgcaagg agtttgagcg agggcacttc
     2281 aaacaatgct gggtggaaca gaagttctct ggctgttggt ctgcgagcag gctcagactg
     2341 caggcacttt tgaatgaact ccctctgtaa tgggtcttct agaagctgga tggcactgct
     2401 gatggcttcc tgtggcacat atgaggactc tccattgccc tgaatctcca gcactgccat
     2461 ctccagtgca cacatgccaa aggagtagat gtccactgct gttgtcacat tagtgacttc
     2521 tccatactct ggtgcaaaga agtgtagatt cttctgctct tctcgacaag tcttcacatg
     2581 attgttgata gtgtcaggag ccacagagcc aatcttgatg agtccgttgt gctggatgaa
     2641 gatggtgtca caggtcaggt tcccatggat gatggggggg tcacaggagt gcaggtagct
     2701 tagggcagag aggatttgtg tgcaccaacg cttccatgcc ttttcattca tcgtcttgtg
     2761 gttctttttg gtcttcttca gaaattgctt cagactccca gatgacatgt attctgtgat
     2821 aaaaatgacc ctggccttgt tctctttaat gtcagcccaa tatttgtgaa acttaacaat
     2881 gttaagatgc tccaattgaa tcagattatc aaacacagca cgaacctttt cctcctgcag
     2941 cttgtagttc ttgcgttcag agaactgtac ctcattccac acaacctcta caccttcctc
     3001 tgtatccatg gccaggtatg cactgtcaat acctggtaca ttccgttgat tcacctcttc
     3061 tcgcctcttc tgccagcgcc cacagggcga ctcttccaaa atctcagact catcttcact
     3121 ttcttcttct tcctctgggg aagcagctga ggttgtggag gtcacaggag gtgacactga
     3181 tgtcaggcca ggagctgaag atgaggattc tacctttggg tctgagccac tgctaagtac
     3241 tgtctgggac tccccctccg acttgtcatc gtcgtccttg tagtccatgc tagcggatct
     3301 gacggttcac taaaccagct ctgcttatat agacctccca ccgtacacgc ctaccgccca
     3361 tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc
     3421 aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc
     3481 gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta
     3541 atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc
     3601 aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatagtcca
     3661 ttttaaaaca taattttaaa actgcaaact acccaagaaa ttattacttt ctacgtcacg
     3721 tattttgtac taatatcttt gtgtttacag tcaaattaat tccaattatc tctctaacag
     3781 ccttgtatcg tatatgcaaa tatgaaggaa tcatgggaaa taggccctcg gtgaaggggg
     3841 cggccgctcg aggctagtct cgtgatcgat accgtcgaga tccgttcact aatcgaatgg
     3901 atctgtctct gtctctctct ccaccttctt cttctattcc ttcgggcctg tcgggtcccc
     3961 tcggggttgg gaggtgggtc tgaaacgata atggtgaata tccctgccta actctattca
     4021 ctatagaaag tacagcaaaa actattctta aacctaccaa gcctcctact atcattatga
     4081 ataattttat ataccacagc caatttgtta tgttaaacca attccacaaa cttgcccatt
     4141 tatctaattc caataattct tgttcattct tttcttgctg gttttgcgat tcttcaatta
     4201 aggagtgtat taagcttgtg taattgttaa tttctctgtc ccactccatc caggtcgtgt
     4261 gattccaaat ctgttccaga gatttattac tccaactagc attccaaggc acagcagtgg
     4321 tgcaaatgag ttttccagag caaccccaaa tccccaggag ctgttgatcc tttaggtatc
     4381 tttccacagc caggattctt gcctggagct gcttgatgcc ccagactgtg agttgcaaca
     4441 gatgctgttg cgcctcaata gccctcagca aattgttctg ctgctgcact ataccagaca
     4501 ataattgtct ggcctgtacc gtcagcgtca ttgacgctgc gcccatagtg cttcctgctg
     4561 ctcccaagaa cccaaggaac aaagctccta ttcccactgc tcttttttct ctctgcacca
     4621 ctcttctctt tgccttggtg ggtgctactc ctaatggttc aatttttact actttatatt
     4681 tatataattc acttctccaa ttgtccctca tatctcctcc tccaggtctg aagatcagcg
     4741 gccgcttgct gtgcggtggt cttacttttg ttttgctctt cctctatctt gtctaaagct
     4801 tccttggtgt cttttatctc tatcctttga tgcacacaat agagggttgc tactgtatta
     4861 tataatgatc taagttcttc tgatcctgtc tgaagggatg gttgtagctg tcccagtatt
     4921 tgtctacagc cttctgatgt ttctaacagg ccaggattaa ctgcgaatcg ttctagctcc
     4981 ctgcttgccc atactatatg ttttaattta tattttttct ttccccctgg ccttaaccga
     5041 attttttccc atcgcgatct aattctcccc cgcttaatac tgacgctctc gcacccatct
     5101 ctctccttct agcctccgct agtcaaaatt tttggcgtac tcaccagtcg ccgcccctcg
     5161 cctcttgccg tgcgcgcttc agcaagccga gtcctgcgtc gagagagctc ctctggtttc
     5221 cctttcgctt tcaagtccct gttcgggcgc cactgctaga gattttccac actgactaaa
     5281 agggtctgag ggatctctag ttaccagagt cacacaacag acgggcacac actacttgaa
     5341 gcactcaagg caagctttat tgaggcttaa gcagtgggtt ccctagttag ccagagagct
     5401 cccaggctca gatctggtct aaccagagag acccgtttat gtatcgagct aggcacttaa
     5461 atacaatatc tctgcaatgc ggcaattcag tggttcgtcc aatccatgtc agacccgtct
     5521 gttgccttcc taataaggca cgatcgtacc accttacttc caccaatcgg catgcacggt
     5581 gctttttctc tccttgtaag gcatgttgct aactcatcgt taccatgttg caagactaca
     5641 agagtattgc ataagactac attaagcttg cagctccagc ttttgttccc tttagtgagg
     5701 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5761 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5821 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5881 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5941 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6001 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6061 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6121 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6181 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6241 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6301 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6361 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6421 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6481 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6541 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6601 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6661 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6721 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6781 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6841 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6901 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6961 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7021 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7081 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7141 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7201 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7261 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7321 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7381 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7441 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7501 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7561 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7621 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7681 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7741 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7801 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7861 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     7921 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     7981 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     8041 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     8101 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     8161 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     8221 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     8281 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     8341 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     8401 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     8461 tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag
     8521 ggcgaattgg gtacgtccct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     8581 cagaggccga ggcggcctcg gcctctgcat aaat
//
