LOCUS       sequence-371140-        8474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression vector coding for Blasticidin, iRFP and 24X 
            MS2 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 8474)
  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 8474)
  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 - 24XMS2 Lentiviral Vector
FEATURES             Location/Qualifiers
     source          1..8474
                     /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    3653..3686
                     /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    3687..3911
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3900..3917
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3949..4347)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(4357..4410)
                     /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(4411..5358)
                     /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    5361..5386
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    5387..5420
                     /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             5527..5760
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5879..5897)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5965..6069
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6070..6138,6139..6930)
                     /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(6288..6307)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7101..7689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7590..7609
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7843..7860
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8100..8121
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8136..8141,8142..8159,8160..8166)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8174..8190
                     /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     8179..8201
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8198..8214
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8198..8214
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8264..8283
                     /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 aattggcata cattatacga agttatggtg acactataga
     2281 ataggatctg atgaaccctg gaatactgga gcgacgcgtg atcgtcgtcg tttgaagatt
     2341 cgacctggag gcacggctga tgctcgtgct ttcttggcaa taagtaccgt aggatctgat
     2401 gaaccctgga atactggagc gacgcgtgat cgtcgtcgtt tgaagattcg acctggaggc
     2461 acggctgatg ctcgtgcttt cttggcaata agtaccgtag gatctgatga accctggaat
     2521 actggagcga cgcgtgatcg tcgtcgtttg aagattcgac ctggaggcac ggctgatgct
     2581 cgtgctttct tggcaataag taccgtagga tctgatgaac cctggaatac tggagcgacg
     2641 cgtgatcgtc gtcgtttgaa gattcgacct ggaggcacgg ctgatgctcg tgctttcttg
     2701 gcaataagta ccgtaggatc tgatgaaccc tggaatactg gagcgacgcg tgatcgtcgt
     2761 cgtttgaaga ttcgacctgg aggcacggct gatgctcgtg ctttcttggc aataagtacc
     2821 gtaggatctg atgaaccctg gaatactgga gcgacgcgtg atcgtcgtcg tttgaagatt
     2881 cgacctggag gcacggctga tgctcgtgct ttcttggcaa taagtaccgt aggatctgat
     2941 gaaccctgga atactggagc gacgcgtgat cgtcgtcgtt tgaagattcg acctggaggc
     3001 acggctgatg ctcgtgcttt cttggcaata agtaccgtag gatctgatga accctggaat
     3061 actggagcga cgcgtgatcg tcgtcgtttg aagattcgac ctggaggcac ggctgatgct
     3121 cgtgctttct tggcaataag taccgtagga tctgatgaac cctggaatac tggagcgacg
     3181 cgtgatcgtc gtcgtttgaa gattcgacct ggaggcacgg ctgatgctcg tgctttcttg
     3241 gcaataagta ccgtaggatc tgatgaaccc tggaatactg gagcgacgcg tgatcgtcgt
     3301 cgtttgaaga ttcgacctgg aggcacggct gatgctcgtg ctttcttggc aataagtacc
     3361 gtaggatctg atgaaccctg gaatactgga gcgacgcgtg atcgtcgtcg tttgaagatt
     3421 cgacctggag gcacggctga tgctcgtgct ttcttggcaa taagtaccgt aggatctgat
     3481 gaaccctgga atactggagc gacgcgtgat cgtcgtcgtt tgaagattcg acctggaggc
     3541 acggctgatg ctcgtgcttt cttggcaata agtaccgtag gatcctctag aattcagatc
     3601 tatagcttag acccactttg tatacaactc ttgtgtagcg attatggatg gcgaagttcc
     3661 tattctctag aaagtatagg aacttcccat agagcccacc gcatccccag catgcctgct
     3721 attgtcttcc caatcctccc ccttgctgtc ctgccccacc ccacccccca gaatagaatg
     3781 acacctactc agacaatgcg atgcaatttc ctcattttat taggaaagga cagtgggagt
     3841 ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa acaacagatg gctggcaact
     3901 agaaggcaca gtcgaggctg atcagcgggt ttaaacgggc cctctagatc agccctccca
     3961 cacataacca gagggcagca attcacgaat cccaactgcc gtcggctgtc catcactgtc
     4021 cttcactatg gctttgatcc caggatgcag atcgagaagc acctgtcggc accgtccgca
     4081 ggggctcaag atgcccctgt tctcatttcc gatcgcgacg atacaagtca ggttgcctgc
     4141 agccgcagca gcagcagtgc ccagcaccac gagttctgca caaggtcccc cagtaaaatg
     4201 atatacattg acaccagtga agatgcggcc gtcgctagag agagctgcgc tggcgacgct
     4261 gtagtcttca gagatgggga tgctgttgat tgtagccgtt gctctttcaa tgagggtgga
     4321 ttcttcttga gacaaaggct tggccatggc gcgcctaggg ccgggattct cctccacgtc
     4381 accgcatgtt agaagacttc ctctgccctc ctcttccatc acgccgatct gccaggccag
     4441 aacctgggcg actagctcgc aggcttggcg gccatcgaga tcgacgtagt acggcgttcg
     4501 gtgatggcaa acgatcaatc cccacagtcg ctcgccgcgc aaaatcgaga tcgacatcgt
     4561 gccgtgcatg cctatgttgc gcatgaattc cagatggacg ggcgagacgc tgcgcaggat
     4621 ggcgaagcta agatcaatcg gccgcccggt gaccggattg aggtctgggg tgaccggcac
     4681 cggccgataa ttgatatcgg gaatgatccg taccgggttg atggtataga gccgacgggc
     4741 ctgcgccggc acggttgagg caggatagtg caggcctagt tttgactcga cctcggcgca
     4801 ccgatcctct gcgatcactt cgccgctgaa gtcggaggcg aagcgataga tcatcacccg
     4861 atcgaagccg gtaatcttcc gcacctcttg cgccgcggcg gcgcaggcgc tttccaaggt
     4921 ttcggcggcc tgcaggcggc ggatggcgct gttggtgcgg cggaagaacg cctgcggctc
     4981 ggcgacgtcc cgctggggag gctcgagctc gaggaagatg agctgatcat ggcgatgcca
     5041 ggagccgatg aagcctgcgt cctttcgcat cgtgaagccg acagtgatcg gtgctccgac
     5101 ggccgccccg ggctcggcca aggcgatcgt cagacggttg tgcgtctccg agtcgaagac
     5161 atcggccgca gagcggccga tcagggcgcc gatcgccagt ccggtgagtt cgggaaggtt
     5221 gtcgctgccg gcaacgatcg tcatgtcggc ggcgagggcg agcagcagtc catgcggttg
     5281 gatggcaccg gggatatgga tcggctcatc gtcgcaggtc aagaggtcag gctgcctggc
     5341 gacggatcct tcagccatgc cctgtgggag gaagagaaga ggtcaggaag ttcctattct
     5401 ctagaaagta taggaacttc gataaatgcc cagattctgc cgctctgtac acctgagatc
     5461 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     5521 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
     5581 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     5641 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     5701 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     5761 gtagtagttc atgtcatctt attattcagt atctataact tgcaaagaaa tgaatatcag
     5821 agagtgagag gcccgggtta attaaggaaa gggctagatc attcttgaag acgaaagggc
     5881 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     5941 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6001 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6061 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6121 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6181 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6241 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6301 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6361 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6421 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6481 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6541 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6601 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6661 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6721 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6781 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6841 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6901 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6961 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7021 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7081 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7141 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7201 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7261 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7321 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7381 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7441 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7501 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7561 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7621 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7681 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7741 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     7801 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     7861 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     7921 tgcagcaagc tcatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     7981 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     8041 agctccccgt ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     8101 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     8161 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgacatgatt
     8221 acgaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     8281 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc
     8341 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa
     8401 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact
     8461 acaaacttag tagt
//
