LOCUS       Exported                8953 bp ds-DNA     circular SYN 17-NOV-2023
DEFINITION  Aguilera_Uribe_W6Y_7_N1     coverage: 938x.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8953)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jun 12, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8953
                     /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(3601..3624)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3656..3859)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             complement(4466..4510)
                     /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    4695..4928
                     /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    5421..5546
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5593..5773
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(5774..6000)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     promoter        complement(6028..6046)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6067..6083)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6091..6107
                     /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(6115..6145)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6160..6181
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6469..7057)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7228..8088)
                     /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(8089..8193)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8219..8674)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     8816..8832
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8842..8860
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      join(8881..8953,1..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 aattcctaag aggagacggt gacagcggct gagttgaggg tactgttcct
     2041 ggcaaaattg aacttgttca aggtctcttc tagcagagaa gtcaaccggc tctggtcagc
     2101 ctcactaatg aagcccagct gcaccagctc agccgccaac tcggggatat tctcatttgg
     2161 catcaggtca cagctcaggt gccggttcag tttgtcctcc aacttcagca gaagtgtcag
     2221 gtggtgtttg actccctcct ccaccgactc aatgttgcac tgcatcagca ccaccttgcg
     2281 agtctccacc tcagctggtt caggtgtcgg agtcttgaca gaggggggca cgacaggtga
     2341 tgtcacctcc tcctgctgtg gctgctgggg ccgaggcagc ccaaaggctg tcagaggata
     2401 gatcccattc ctgacatctt caaggaattt atctaattcc agagctggtg actgagagta
     2461 caaagtctga actggttctc ttcctggtcc tgcagggatt tcagccagta cggcactagt
     2521 atccatgttt ttggtgatct cctctagagc gttctctggg atcatgtgtt ggtgtcccac
     2581 aatgcagtgg gccgcaagga gtttgagcga gggcacttca aacaatgctg ggtggaacag
     2641 aagttctctg gctgttggtc tgcgagcagg ctcagactgc aggcactttt gaatgaactc
     2701 cctctgtaat gggtcttcta gaagctggat ggcactgctg atggcttcct gtggcacata
     2761 tgaggactct ccattgccct gaatctccag cactgccatc tccagtgcac acatgccaaa
     2821 ggagtagatg tccactgctg ttgtcacatt agtgacttct ccatactctg gtgcaaagaa
     2881 gtgtagattc ttctgctctt ctcgacaagt cttcacatga ttgttgatag tgtcaggagc
     2941 cacagagcca atcttgatga gtccgttgtg ctggatgaag atggtgtcac aggtcaggtt
     3001 cccatggatg atgggggggt cacaggagtg caggtagctt agggcagaga ggatttgtgt
     3061 gcaccaacgc ttccatgcct tttcattcat cgtcttgtgg ttctttttgg tcttcttcag
     3121 aaattgcttc agactcccag atgacatgta ttctgtgata aaaatgaccc tggccttgtt
     3181 ctctttaatg tcagcccaat atttgtgaaa cttaacaatg ttaagatgct ccaattgaat
     3241 cagattatca aacacagcac gaaccttttc ctcctgcagc ttgtagttct tgcgttcaga
     3301 gaactgtacc tcattccaca caacctctac accttcctct gtatccatgg ccaggtatgc
     3361 actgtcaata cctggtacat tccgttgatt cacctcttct cgcctcttct gccagcgccc
     3421 acagggcgac tcttccaaaa tctcagactc atcttcactt tcttcttctt cctctgggga
     3481 agcagctgag gttgtggagg tcacaggagg tgacactgat gtcaggccag gagctgaaga
     3541 tgaggattct acctttgggt ctgagccact gctaagtact gtctgggact ccccctccga
     3601 cttgtcatcg tcgtccttgt agtccatgct agcggatctg acggttcact aaaccagctc
     3661 tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt
     3721 tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt
     3781 caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt
     3841 actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag
     3901 taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
     3961 tgacgtcaat agggggcgta cttggcatat gatagtccat tttaaaacat aattttaaaa
     4021 ctgcaaacta cccaagaaat tattactttc tacgtcacgt attttgtact aatatctttg
     4081 tgtttacagt caaattaatt ccaattatct ctctaacagc cttgtatcgt atatgcaaat
     4141 atgaaggaat catgggaaat aggccctcgg tgaagggggc ggccgctcga ggctagtctc
     4201 gtgatcgata ccgtcgagat ccgttcacta atcgaatgga tctgtctctg tctctctctc
     4261 caccttcttc ttctattcct tcgggcctgt cgggtcccct cggggttggg aggtgggtct
     4321 gaaacgataa tggtgaatat ccctgcctaa ctctattcac tatagaaagt acagcaaaaa
     4381 ctattcttaa acctaccaag cctcctacta tcattatgaa taattttata taccacagcc
     4441 aatttgttat gttaaaccaa ttccacaaac ttgcccattt atctaattcc aataattctt
     4501 gttcattctt ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt aagcttgtgt
     4561 aattgttaat ttctctgtcc cactccatcc aggtcgtgtg attccaaatc tgttccagag
     4621 atttattact ccaactagca ttccaaggca cagcagtggt gcaaatgagt tttccagagc
     4681 aaccccaaat ccccaggagc tgttgatcct ttaggtatct ttccacagcc aggattcttg
     4741 cctggagctg cttgatgccc cagactgtga gttgcaacag atgctgttgc gcctcaatag
     4801 ccctcagcaa attgttctgc tgctgcacta taccagacaa taattgtctg gcctgtaccg
     4861 tcagcgtcat tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac ccaaggaaca
     4921 aagctcctat tcccactgct cttttttctc tctgcaccac tcttctcttt gccttggtgg
     4981 gtgctactcc taatggttca atttttacta ctttatattt atataattca cttctccaat
     5041 tgtccctcat atctcctcct ccaggtctga agatcagcgg ccgcttgctg tgcggtggtc
     5101 ttacttttgt tttgctcttc ctctatcttg tctaaagctt ccttggtgtc ttttatctct
     5161 atcctttgat gcacacaata gagggttgct actgtattat ataatgatct aagttcttct
     5221 gatcctgtct gaagggatgg ttgtagctgt cccagtattt gtctacagcc ttctgatgtt
     5281 tctaacaggc caggattaac tgcgaatcgt tctagctccc tgcttgccca tactatatgt
     5341 tttaatttat attttttctt tccccctggc cttaaccgaa ttttttccca tcgcgatcta
     5401 attctccccc gcttaatact gacgctctcg cacccatctc tctccttcta gcctccgcta
     5461 gtcaaaattt ttggcgtact caccagtcgc cgcccctcgc ctcttgccgt gcgcgcttca
     5521 gcaagccgag tcctgcgtcg agagagctcc tctggtttcc ctttcgcttt caagtccctg
     5581 ttcgggcgcc actgctagag attttccaca ctgactaaaa gggtctgagg gatctctagt
     5641 taccagagtc acacaacaga cgggcacaca ctacttgaag cactcaaggc aagctttatt
     5701 gaggcttaag cagtgggttc cctagttagc cagagagctc ccaggctcag atctggtcta
     5761 accagagaga cccgtttatg tatcgagcta ggcacttaaa tacaatatct ctgcaatgcg
     5821 gcaattcagt ggttcgtcca atccatgtca gacccgtctg ttgccttcct aataaggcac
     5881 gatcgtacca ccttacttcc accaatcggc atgcacggtg ctttttctct ccttgtaagg
     5941 catgttgcta actcatcgtt accatgttgc aagactacaa gagtattgca taagactaca
     6001 ttaagcttgc agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     6061 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6121 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     6181 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     6241 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6301 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6361 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6421 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6481 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6541 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6601 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6661 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6721 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6781 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6841 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6901 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6961 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7021 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7081 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7141 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7201 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7261 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7321 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7381 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7441 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7501 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7561 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7621 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7681 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7741 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7801 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7861 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7921 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7981 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8041 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8101 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8161 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     8221 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     8281 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     8341 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     8401 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     8461 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     8521 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     8581 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     8641 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac
     8701 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     8761 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     8821 acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg tacgtcccta
     8881 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     8941 cctctgcata aat
//
