LOCUS       Exported                7691 bp ds-DNA     circular SYN 10-JUN-2024
DEFINITION  Aguilera_Uribe_W6Y_11_N5     coverage: 764x.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7691)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jun 12, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7691
                     /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"
     CDS             complement(2000..2023)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(2024..2362)
                     /codon_start=1
                     /label=NRBP1 C-term
                     /translation="PVVPPSVKTPTPEPAEVETRKVVLMQCNIESVEEGVKHHLTLLLK
                     LEDKLNRHLSCDLMPNENIPELAAELVQLGFISEADQSRLTSLLEETLNKFNFARNSTL
                     NSAAVTVSS"
     promoter        complement(2394..2597)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             complement(3204..3248)
                     /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    3433..3666
                     /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    4159..4284
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4331..4511
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(4512..4738)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     promoter        complement(4766..4784)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    4792..4811
                     /label=N terminal HA- R 3' Gibson
     misc_feature    4792..4811
                     /label=C terminal HA- R 3' Gibson
     primer_bind     complement(4805..4821)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4829..4845
                     /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(4853..4883)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4898..4919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5207..5795)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5966..6826)
                     /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(6827..6931)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6957..7412)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     misc_feature    7552..7571
                     /label=N terminal HA- L 5' Gibson
     misc_feature    7552..7571
                     /label=C terminal HA-L 5' Gibson
     primer_bind     7554..7570
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7580..7598
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      join(7619..7691,1..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 gaattcctac ttgtcatcgt cgtccttgta gtcagaggag acggtgacag
     2041 cggctgagtt gagggtactg ttcctggcaa aattgaactt gttcaaggtc tcttctagca
     2101 gagaagtcaa ccggctctgg tcagcctcac taatgaagcc cagctgcacc agctcagccg
     2161 ccaactcggg gatattctca tttggcatca ggtcacagct caggtgccgg ttcagtttgt
     2221 cctccaactt cagcagaagt gtcaggtggt gtttgactcc ctcctccacc gactcaatgt
     2281 tgcactgcat cagcaccacc ttgcgagtct ccacctcagc tggttcaggt gtcggagtct
     2341 tgacagaggg gggcacgaca ggcatgctag cggatctgac ggttcactaa accagctctg
     2401 cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg
     2461 ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca
     2521 atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac
     2581 tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
     2641 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
     2701 acgtcaatag ggggcgtact tggcatatga tagtccattt taaaacataa ttttaaaact
     2761 gcaaactacc caagaaatta ttactttcta cgtcacgtat tttgtactaa tatctttgtg
     2821 tttacagtca aattaattcc aattatctct ctaacagcct tgtatcgtat atgcaaatat
     2881 gaaggaatca tgggaaatag gccctcggtg aagggggcgg ccgctcgagg ctagtctcgt
     2941 gatcgatacc gtcgagatcc gttcactaat cgaatggatc tgtctctgtc tctctctcca
     3001 ccttcttctt ctattccttc gggcctgtcg ggtcccctcg gggttgggag gtgggtctga
     3061 aacgataatg gtgaatatcc ctgcctaact ctattcacta tagaaagtac agcaaaaact
     3121 attcttaaac ctaccaagcc tcctactatc attatgaata attttatata ccacagccaa
     3181 tttgttatgt taaaccaatt ccacaaactt gcccatttat ctaattccaa taattcttgt
     3241 tcattctttt cttgctggtt ttgcgattct tcaattaagg agtgtattaa gcttgtgtaa
     3301 ttgttaattt ctctgtccca ctccatccag gtcgtgtgat tccaaatctg ttccagagat
     3361 ttattactcc aactagcatt ccaaggcaca gcagtggtgc aaatgagttt tccagagcaa
     3421 ccccaaatcc ccaggagctg ttgatccttt aggtatcttt ccacagccag gattcttgcc
     3481 tggagctgct tgatgcccca gactgtgagt tgcaacagat gctgttgcgc ctcaatagcc
     3541 ctcagcaaat tgttctgctg ctgcactata ccagacaata attgtctggc ctgtaccgtc
     3601 agcgtcattg acgctgcgcc catagtgctt cctgctgctc ccaagaaccc aaggaacaaa
     3661 gctcctattc ccactgctct tttttctctc tgcaccactc ttctctttgc cttggtgggt
     3721 gctactccta atggttcaat ttttactact ttatatttat ataattcact tctccaattg
     3781 tccctcatat ctcctcctcc aggtctgaag atcagcggcc gcttgctgtg cggtggtctt
     3841 acttttgttt tgctcttcct ctatcttgtc taaagcttcc ttggtgtctt ttatctctat
     3901 cctttgatgc acacaataga gggttgctac tgtattatat aatgatctaa gttcttctga
     3961 tcctgtctga agggatggtt gtagctgtcc cagtatttgt ctacagcctt ctgatgtttc
     4021 taacaggcca ggattaactg cgaatcgttc tagctccctg cttgcccata ctatatgttt
     4081 taatttatat tttttctttc cccctggcct taaccgaatt ttttcccatc gcgatctaat
     4141 tctcccccgc ttaatactga cgctctcgca cccatctctc tccttctagc ctccgctagt
     4201 caaaattttt ggcgtactca ccagtcgccg cccctcgcct cttgccgtgc gcgcttcagc
     4261 aagccgagtc ctgcgtcgag agagctcctc tggtttccct ttcgctttca agtccctgtt
     4321 cgggcgccac tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta
     4381 ccagagtcac acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga
     4441 ggcttaagca gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac
     4501 cagagagacc cgtttatgta tcgagctagg cacttaaata caatatctct gcaatgcggc
     4561 aattcagtgg ttcgtccaat ccatgtcaga cccgtctgtt gccttcctaa taaggcacga
     4621 tcgtaccacc ttacttccac caatcggcat gcacggtgct ttttctctcc ttgtaaggca
     4681 tgttgctaac tcatcgttac catgttgcaa gactacaaga gtattgcata agactacatt
     4741 aagcttgcag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat
     4801 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     4861 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     4921 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     4981 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5041 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5101 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5161 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5221 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5281 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5341 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5401 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5461 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5521 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5581 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5641 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5701 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5761 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5821 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5881 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5941 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6001 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6061 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6121 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6181 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6241 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6301 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6361 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6421 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6481 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6541 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6601 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6661 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6721 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6781 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6841 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6901 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
     6961 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     7021 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     7081 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     7141 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     7201 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     7261 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     7321 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     7381 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg
     7441 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     7501 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     7561 gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta cgtccctagg
     7621 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
     7681 tctgcataaa t
//
