LOCUS       sequence-343727-        8602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression vector coding for Blasticidin, iRFP and 24X 
            PP7 hairpins. All inserts are cloned in on the -strand to allow for 
            splicing in when targeted into an intron of transcribed genes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8602)
  AUTHORS   Wan Y, Anastasakis DG, Rodriguez J, Palangat M, Gudla P, Zaki G, 
            Tandon M, Pegoraro G, Chow CC, Hafner M, Larson DR
  TITLE     Dynamic imaging of nascent RNA reveals general principles of 
            transcription dynamics and stochastic splice site selection.
  JOURNAL   Cell. 2021 May 27;184(11):2878-2895.e20. doi: 
            10.1016/j.cell.2021.04.012. Epub 2021 May 11.
  PUBMED    33979654
REFERENCE   2  (bases 1 to 8602)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2021.04.012"; journalName: "Cell"; date: 
            "2021-05-27- 27"; volume: "184"; issue: "11"; pages: "2878-2895.e20"
COMMENT     Sequence Label: Gene Trap - 24XPP7 Lentiviral Vector
FEATURES             Location/Qualifiers
     source          1..8602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    3781..3814
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     polyA_signal    3815..4039
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4028..4045
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(4077..4475)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(4485..4538)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(4539..5486)
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     misc_feature    5489..5514
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    5515..5548
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     LTR             5655..5888
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6007..6025)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6093..6197
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6198..6266,6267..7058)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6416..6435)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7229..7817
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7718..7737
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7971..7988
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8228..8249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8264..8269,8270..8287,8288..8294)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8302..8318
                     /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)."
     primer_bind     8307..8329
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8326..8342
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8326..8342
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8392..8411
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtacc
     2221 ggtgctagct gatatctgtc aattgacgcg ttctcgagat ctaatgaacc cgggaatact
     2281 gcagccagcg agcccatatg ctctgctggt ttctagagtc gacctgcagg gagcgacgcc
     2341 atatcgtctg ctcctttcta ggcaattagg taccttagga tctaatgaac ccgggaatac
     2401 tgcagccagc gagcccatat gctctgctgg tttctagagt cgacctgcag ggagcgacgc
     2461 catatcgtct gctcctttct aggcaattag gtaccttagg atctaatgaa cccgggaata
     2521 ctgcagccag cgagcccata tgctctgctg gtttctagag tcgacctgca gggagcgacg
     2581 ccatatcgtc tgctcctttc taggcaatta ggtaccttag gatctaatga acccgggaat
     2641 actgcagcca gcgagcccat atgctctgct ggtttctaga gtcgacctgc agggagcgac
     2701 gccatatcgt ctgctccttt ctaggcaatt aggtacctta ggatctaatg aacccgggaa
     2761 tactgcagcc agcgagccca tatgctctgc tggtttctag agtcgacctg cagggagcga
     2821 cgccatatcg tctgctcctt tctaggcaat taggtacctt aggatctaat gaacccggga
     2881 atactgcagc cagcgagccc atatgctctg ctggtttcta gagtcgacct gcagggagcg
     2941 acgccatatc gtctgctcct ttctaggcaa ttaggtacct taggatctaa tgaacccggg
     3001 aatactgcag ccagcgagcc catatgctct gctggtttct agagtcgacc tgcagggagc
     3061 gacgccatat cgtctgctcc tttctaggca attaggtacc ttaggatcta atgaacccgg
     3121 gaatactgca gccagcgagc ccatatgctc tgctggtttc tagagtcgac ctgcagggag
     3181 cgacgccata tcgtctgctc ctttctaggc aattaggtac cttaggatct aatgaacccg
     3241 ggaatactgc agccagcgag cccatatgct ctgctggttt ctagagtcga cctgcaggga
     3301 gcgacgccat atcgtctgct cctttctagg caattaggta ccttaggatc taatgaaccc
     3361 gggaatactg cagccagcga gcccatatgc tctgctggtt tctagagtcg acctgcaggg
     3421 agcgacgcca tatcgtctgc tcctttctag gcaattaggt accttaggat ctaatgaacc
     3481 cgggaatact gcagccagcg agcccatatg ctctgctggt ttctagagtc gacctgcagg
     3541 gagcgacgcc atatcgtctg ctcctttcta ggcaattagg taccttagga tctaatgaac
     3601 ccgggaatac tgcagccagc gagcccatat gctctgctgg tttctagagt cgacctgcag
     3661 ggagcgacgc catatcgtct gctcctttct aggcaattag gtaccttagg atctcgagta
     3721 tcgatacatt acactttaac ccactttgta tacaactctt gtgtagcgat tatggatggc
     3781 gaagttccta ttctctagaa agtataggaa cttcccatag agcccaccgc atccccagca
     3841 tgcctgctat tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga
     3901 atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca
     3961 gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc
     4021 tggcaactag aaggcacagt cgaggctgat cagcgggttt aaacgggccc tctagatcag
     4081 ccctcccaca cataaccaga gggcagcaat tcacgaatcc caactgccgt cggctgtcca
     4141 tcactgtcct tcactatggc tttgatccca ggatgcagat cgagaagcac ctgtcggcac
     4201 cgtccgcagg ggctcaagat gcccctgttc tcatttccga tcgcgacgat acaagtcagg
     4261 ttgcctgcag ccgcagcagc agcagtgccc agcaccacga gttctgcaca aggtccccca
     4321 gtaaaatgat atacattgac accagtgaag atgcggccgt cgctagagag agctgcgctg
     4381 gcgacgctgt agtcttcaga gatggggatg ctgttgattg tagccgttgc tctttcaatg
     4441 agggtggatt cttcttgaga caaaggcttg gccatggcgc gcctagggcc gggattctcc
     4501 tccacgtcac cgcatgttag aagacttcct ctgccctcct cttccatcac gccgatctgc
     4561 caggccagaa cctgggcgac tagctcgcag gcttggcggc catcgagatc gacgtagtac
     4621 ggcgttcggt gatggcaaac gatcaatccc cacagtcgct cgccgcgcaa aatcgagatc
     4681 gacatcgtgc cgtgcatgcc tatgttgcgc atgaattcca gatggacggg cgagacgctg
     4741 cgcaggatgg cgaagctaag atcaatcggc cgcccggtga ccggattgag gtctggggtg
     4801 accggcaccg gccgataatt gatatcggga atgatccgta ccgggttgat ggtatagagc
     4861 cgacgggcct gcgccggcac ggttgaggca ggatagtgca ggcctagttt tgactcgacc
     4921 tcggcgcacc gatcctctgc gatcacttcg ccgctgaagt cggaggcgaa gcgatagatc
     4981 atcacccgat cgaagccggt aatcttccgc acctcttgcg ccgcggcggc gcaggcgctt
     5041 tccaaggttt cggcggcctg caggcggcgg atggcgctgt tggtgcggcg gaagaacgcc
     5101 tgcggctcgg cgacgtcccg ctggggaggc tcgagctcga ggaagatgag ctgatcatgg
     5161 cgatgccagg agccgatgaa gcctgcgtcc tttcgcatcg tgaagccgac agtgatcggt
     5221 gctccgacgg ccgccccggg ctcggccaag gcgatcgtca gacggttgtg cgtctccgag
     5281 tcgaagacat cggccgcaga gcggccgatc agggcgccga tcgccagtcc ggtgagttcg
     5341 ggaaggttgt cgctgccggc aacgatcgtc atgtcggcgg cgagggcgag cagcagtcca
     5401 tgcggttgga tggcaccggg gatatggatc ggctcatcgt cgcaggtcaa gaggtcaggc
     5461 tgcctggcga cggatccttc agccatgccc tgtgggagga agagaagagg tcaggaagtt
     5521 cctattctct agaaagtata ggaacttcga taaatgccca gattctgccg ctctgtacac
     5581 ctgagatcct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     5641 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc
     5701 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     5761 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     5821 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     5881 ctctagcagt agtagttcat gtcatcttat tattcagtat ctataacttg caaagaaatg
     5941 aatatcagag agtgagaggc ccgggttaat taaggaaagg gctagatcat tcttgaagac
     6001 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     6061 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     6121 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     6181 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     6241 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     6301 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     6361 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     6421 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     6481 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     6541 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6601 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6661 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6721 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6781 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6841 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6901 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6961 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     7021 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     7081 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     7141 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     7201 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     7261 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     7321 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     7381 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     7441 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     7501 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7561 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7621 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7681 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7741 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7801 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7861 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     7921 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     7981 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     8041 ttcattaatg cagcaagctc atggctgact aatttttttt atttatgcag aggccgaggc
     8101 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     8161 ttgcaaaaag ctccccgtgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     8221 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     8281 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     8341 acatgattac gaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     8401 ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa
     8461 atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt
     8521 tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa
     8581 tatgtactac aaacttagta gt
//
