LOCUS       sequence-253024-        6438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This vector is used for cloning a gene driven by 4xnrUAS, to be 
            co-expressed with mCerulean3 through P2A self-cleaving peptides..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6438)
  AUTHORS   Gu Q, Yang X, Lv J, Zhang J, Xia B, Kim JD, Wang R, Xiong F, Meng S,
            Clements TP, Tandon B, Wagner DS, Diaz MF, Wenzel PL, Miller YI, 
            Traver D, Cooke JP, Li W, Zon LI, Chen K, Bai Y, Fang L
  TITLE     AIBP-mediated cholesterol efflux instructs hematopoietic stem and 
            progenitor cell fate.
  JOURNAL   Science. 2019 Mar 8;363(6431):1085-1088. doi: 
            10.1126/science.aav1749. Epub 2019 Jan 31.
  PUBMED    30705153
REFERENCE   2  (bases 1 to 6438)
  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: 
            "10.1126/science.aav1749"; journalName: "Science"; date: "2019-03-8-
            8"; volume: "363"; issue: "6431"; pages: "1085-1088"
COMMENT     Sequence Label: pBT2-4xUAS:P2A-mCerulean3
FEATURES             Location/Qualifiers
     source          1..6438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(1..12,13..200)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     1006..1022
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1007..1023
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1327..1383
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(1390..1392,1393..1395,1396..2109)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIK
                     ANFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1435..1456)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1696..1715)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2043..2064
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2148..2282
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2172..2191)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2226..2245
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(3403..3565,3566..3577)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     promoter        complement(3592..3610)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3617..3634)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3617..3633)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3626..3648)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3775..4230
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3862..3881)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4072..4093
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4256..4360
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4361..4429,4430..5221)
                     /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     complement(4579..4598)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5392..5980
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5881..5900
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6134..6151
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6268..6289
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6304..6309,6310..6327,6328..6334)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6342..6358
                     /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     6347..6369
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6366..6382
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6366..6382
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6403..6421
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cagaggtgta aagtacttga gtaattttac ttgattactg tacttaagta ttatttttgg
       61 ggatttttac tttacttgag tacaattaaa aatcaatact tttactttta cttaattaca
      121 tttttttaga aaaaaaagta ctttttactc cttacaattt tatttacagt caaaaagtac
      181 ttattttttg gagatcactt cattctattt tcccttgcta ttaccaaacc aattgaattg
      241 cgctgatgcc cagtttaatt taaatgttat ttattctgcc tatgaaaatc gttttcacat
      301 tatatgaaat tggtcagaca tgttcattgg tcctttggaa gtgacgtcat gtcacatcta
      361 ttaccacaat gcacagcacc ttgacctgga aattagggaa attataacag tcaatcagtg
      421 gaagaaaatg gaggaagtat gtgattcatc agcagctgcg agcagcacag tccaaaatca
      481 gccacaggat caagagcacc cgtggccgta tcttcgcgaa ttcttttctt taagtggtgt
      541 aaataaagat tcattcaaga tgaaatgtgt cctctgtctc ccgcgggccc tacgaagtta
      601 tagatcctct agcgataagc ttgatatcga attcgagttg gcgcgcctgt cattctaaat
      661 ctctctttca gcctaaagct ttttccccgt atccccccag gtgtctgcag gctcaaagag
      721 cagcgagaag cgttcagagg aaagcgatcc cgtgccacct tccccgtgcc cgggctgtcc
      781 ccgcacgctg ccggctcggg gatgcggggg gagcgccgga ccggagcgga gccccgggcg
      841 gctcgctgct gccccctagc gggggaggga cgtaattaca tccctggggg ctttgggggg
      901 gggctgtccc cgtgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct
      961 ccgatagact gcgtcgcccg ggtacccgtg tatccaataa acccctcgag gtcgacggta
     1021 tcgataagct tgatatcgaa ttcctgcagc ccgggggatc acgcggccat caagcttagg
     1081 cctccacggt ggcttctaat ccgtgagtcc tagcgggtga cagccctccg tcttcacagg
     1141 cggaggagag tcttccgtag ggttcctcgg agtactgtcc tccgacgcgt gcaagggtcg
     1201 actctagagg gtatataatg gatcccatcg cgtctcagcc tcactttgag ctcctccaca
     1261 cgaattccct cgacctcgaa gacgcgtgga tccaccggtg gatccctcga gcccgggtct
     1321 agaggagcta ctaacttcag cctgctgaag caggctggag acgtggagga gaaccctgga
     1381 cctagatcca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1441 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1501 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1561 tggcccaccc tcgtgaccac cctgagctgg ggcgtgcagt gcttcgcccg ctaccccgac
     1621 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1681 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1741 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1801 ctggggcaca agctggagta caacgccatc cacggcaacg tctatatcac cgccgacaag
     1861 cagaagaacg gcatcaaggc caacttcggc ctcaactgca acatcgagga cggcagcgtg
     1921 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1981 gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat
     2041 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2101 tacaagtaat cgattctagc tagaactata gtgagtcgta ttacgtagat ccagacatga
     2161 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     2221 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2281 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     2341 tttaattcgc ggccgcggcg actagtgggt ttattggata cacgggtacc cgggcgacgc
     2401 agtctatcgg aggactggcg cgccgagtga ggggttgtgg gctcttttat tgagctcacg
     2461 gggacagccc ccccccaaag cccccaggga tgtaattacg tccctccccc gctagggggc
     2521 agcagcgagc cgcccggggc tccgctccgg tccggcgctc cccccgcatc cccgagccgg
     2581 cagcgtgcgg ggacagcccg ggcacgggga aggtggcacg ggatcgcttt cctctgaacg
     2641 cttctcgctg ctctttgagc ctgcagacac ctggggggat acggggaaaa agctttaggc
     2701 tgaaagagag atttagaatg acaggcgcgc ctgggagctc acggggacag ccccctccca
     2761 aagcccccag ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg
     2821 ggctccgctc cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc
     2881 ccgggcacgg ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg
     2941 agcctgcaga cacctggggg gatacgggga aaaagcttta ggctgaaaga gagatttaga
     3001 atgacaggcg cgccaactcg aattcgatat caagcttatc gctagaggat ctataacttc
     3061 gtaagatctg tctctgctca cgtttcctgc tatttgcagc ctctctatca agactaatac
     3121 acctcttccc gcatcggctg cctgtgagag gcttttcagc actgcaggat tgcttttcag
     3181 ccccaaaaga gctaggcttg acactaacaa ttttgagaat cagcttctac tgaagttaaa
     3241 tctgaggttt tacaactttg agtagcgtgt actggcatta gattgtctgt cttatagttt
     3301 gataattaaa tacaaacagt tctaaagcag gataaaacct tgtatgcatt tcatttaatg
     3361 ttttttgaga ttaaaagctt aaacaagaat ctctagtttt ctttcttgct tttactttta
     3421 cttccttaat actcaagtac aattttaatg gagtactttt ttacttttac tcaagtaaga
     3481 ttctagccag atacttttac ttttaattga gtaaaatttt ccctaagtac ttgtactttc
     3541 acttgagtaa aatttttgag tactttttac acctctggag ctccaattcg ccctatagtg
     3601 agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     3661 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     3721 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     3781 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3841 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3901 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3961 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4021 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4081 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4141 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4201 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     4261 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4321 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4381 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4441 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4501 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4561 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4621 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4681 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4741 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4801 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4861 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4921 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4981 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5041 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5101 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5161 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5221 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5281 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5341 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5401 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5461 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5521 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5581 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5641 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5701 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5761 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5821 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5881 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5941 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6001 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6061 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     6121 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     6181 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     6241 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     6301 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     6361 tcacacagga aacagctatg accatgatta cgccaagctc gaaattaacc ctcactaaag
     6421 ggaacaaaag ctggtacc
//
