LOCUS       sequence-324348-        8036 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of mIFP with multiple cloning site upstream 
            the pEF promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8036)
  AUTHORS   Tycko J, DelRosso N, Hess GT, Aradhana, Banerjee A, Mukund A, Van 
            MV, Ego BK, Yao D, Spees K, Suzuki P, Marinov GK, Kundaje A, Bassik 
            MC, Bintu L
  TITLE     High-Throughput Discovery and Characterization of Human 
            Transcriptional Effectors.
  JOURNAL   Cell. 2020 Dec 9. pii: S0092-8674(20)31541-5. doi: 
            10.1016/j.cell.2020.11.024.
  PUBMED    33326746
REFERENCE   2  (bases 1 to 8036)
  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"; journalName: "Cell. 2020 
            Dec 9. pii: S0092-8674(20)31541-5. doi: 10.1016/j.cell.2020.11.024."
COMMENT     Sequence Label: pJT040 lenti MCS pEF mIFP WPRE
FEATURES             Location/Qualifiers
     source          1..8036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(347..1138,1139..1207))
                     /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"
     primer_bind     970..989
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1208..1312)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1387..1406)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1578..1957
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1959..2157
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2112..2132
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2175..2355
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2402..2527
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3024..3257
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3442..3486
                     /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    3784..3901
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4024..4235
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          4236..5174
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5131..5151
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             5194..6153
                     /codon_start=1
                     /product="naturally monomeric infrared fluorescent protein 
                     derived from a truncated bacteriophytochrome (Yu et al., 
                     2015)"
                     /label=mIFP
                     /translation="MSVPLTTSAFGHAFLANCEREQIHLAGSIQPHGILLAVKEPDNVV
                     IQASINAAEFLNTNSVVGRPLRDLGGDLPLQILPHLNGPLHLAPMTLRCTVGSPPRRVD
                     CTIHRPSNGGLIVELEPATKTTNIAPALDGAFHRITSSSSLMGLCDETATIIREITGYD
                     RVMVVRFDEEGNGEILSERRRADLEAFLGNRYPASTIPQIARRLYEHNRVRLLVDVNYT
                     PVPLQPRISPLNGRDLDMSLSCLRSMSPIHQKYMQDMGVGATLVCSLMVSGRLWGLIAC
                     HHYEPRFVPFHIRAAGEALAETCAIRIATLESFAQSQSK"
     protein_bind    complement(6176..6209)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6265..6853
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6318..6338)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6736..6747)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6856..6872)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6857..6873)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7382..7562
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(join(7624..8036,1..176))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7704..7723)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
       61 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
      121 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
      181 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
      241 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
      301 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
      361 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
      421 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
      481 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
      541 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
      601 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
      661 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
      721 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
      781 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
      841 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
      901 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
      961 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     1021 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     1081 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     1141 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     1201 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     1261 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     1321 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     1381 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     1441 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     1501 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
     1561 ggccagatat acgcgttgac attgattatt gactagttat taatagtaat caattacggg
     1621 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     1681 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     1741 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     1801 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     1861 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     1921 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     1981 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     2041 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     2101 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagcgc
     2161 gttttgcctg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     2221 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     2281 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     2341 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
     2401 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
     2461 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
     2521 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
     2581 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     2641 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     2701 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     2761 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     2821 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg gccgctgatc
     2881 ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa
     2941 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag
     3001 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga
     3061 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt
     3121 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa
     3181 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta
     3241 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct
     3301 gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc
     3361 tggatggagt gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa
     3421 gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca
     3481 agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg
     3541 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga
     3601 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc
     3661 gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag atccattcga
     3721 ttagtgaacg gatcggcact gcgtgcgcca attctgcaga caaatggcag tattcatcca
     3781 caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     3841 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     3901 tcgggtttat tacagggaca gcagagatcc agtttggtta gtaccgggcc cgctctagag
     3961 atcccatttg actcaggcgg ccgcgtggat aaccgtatta ccgccatgca tgtgcccgtc
     4021 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
     4081 gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
     4141 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
     4201 ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg
     4261 ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc acctggctgc
     4321 agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt
     4381 gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc
     4441 gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag
     4501 ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa
     4561 atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg
     4621 cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa
     4681 tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt
     4741 gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa
     4801 gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag
     4861 agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt
     4921 catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc tcgcgctttt
     4981 ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg
     5041 agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg
     5101 ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt
     5161 ttcttccatt tcaggtgtcg tgagctagcc accatgtcgg taccgctgac tacctcagca
     5221 ttcggccacg cgtttctggc taactgtgaa cgcgagcaga tccacctggc gggctccatt
     5281 cagccgcacg gtatcctgct ggctgtgaaa gagccggaca acgtggtgat ccaggcttct
     5341 attaacgctg cggagttcct gaacaccaac tctgttgttg gccgtccgct gcgtgacctg
     5401 ggcggcgatc tgcctttgca gatcctgccg cacctgaacg gcccgctgca cctggctccg
     5461 atgaccctgc gttgtaccgt gggttctccg ccgcgtcgtg tggactgtac cattcatcgt
     5521 ccgtctaacg gcggcctgat cgtagaactg gaaccagcaa ccaagaccac taacattgcg
     5581 ccggctctgg acggtgcgtt tcatcgtatc acttcttcat cctccctgat gggcctgtgt
     5641 gacgaaaccg cgactattat ccgtgagatt actggctacg accgtgtgat ggtagtacgt
     5701 ttcgatgaag agggtaatgg cgaaattctg tccgaacgtc gtcgtgcgga cctggaagcg
     5761 ttcctgggta accgctaccc ggcgtctact attccgcaga tcgctcgtcg cctgtacgaa
     5821 cataaccgtg ttcgcctgct ggtagatgtg aactatactc cggttccgct acagccgcgc
     5881 atcagcccgc tgaacggtcg tgatctggat atgtccctgt cttgcctgcg ctctatgtcc
     5941 ccgatccacc agaaatacat gcaggacatg ggcgttggcg cgaccctggt ttgctctctg
     6001 atggtgtctg gtcgtctgtg gggtctgatc gcttgccacc actacgaacc gcgcttcgtt
     6061 ccgttccaca ttcgcgctgc tggcgaagcg ctggcggaaa cttgtgcgat ccgcatcgcg
     6121 acgctggaga gctttgcaca gtctcagtcc aaataggaat tcgtcgaggg acctaataac
     6181 ttcgtatagc atacattata cgaagttata catgtttaag ggttccggtt ccactaggta
     6241 caattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     6301 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     6361 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     6421 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     6481 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     6541 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     6601 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     6661 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     6721 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     6781 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     6841 gccgcctccc cgcatcgata ccgtcgacct cgatcgagac ctagaaaaac atggagcaat
     6901 cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga
     6961 ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc
     7021 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     7081 ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga
     7141 ttggcagaac tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta
     7201 caagctagta ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg
     7261 cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg
     7321 gaggtttgac agccgcctag catttcatca catggcccga gagctgcatc cggactgtac
     7381 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     7441 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     7501 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     7561 cagcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     7621 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     7681 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     7741 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     7801 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7861 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7921 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7981 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagt
//
