LOCUS       sequence-254848-        6721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This vector is used for cloning a gene driven by 4xnrUAS, to be 
            co-expressed with EGFP through P2A self-cleaving peptides..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6721)
  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 6721)
  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-EGFP
FEATURES             Location/Qualifiers
     source          1..6721
                     /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     1289..1305
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1290..1306
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1610..1666
                     /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(1673..1675,1676..1678,1679..2392)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1673..1675,1676..1678,1679..2389)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1718..1739)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1979..1998)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2326..2347
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2431..2565
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2455..2474)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2509..2528
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(3686..3848,3849..3860)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     promoter        complement(3875..3893)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3900..3917)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3900..3916)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3909..3931)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4058..4513
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4145..4164)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4355..4376
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4539..4643
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4644..4712,4713..5504)
                     /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(4862..4881)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5675..6263
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6164..6183
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6417..6434
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6551..6572
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6587..6592,6593..6610,6611..6617)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6625..6641
                     /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     6630..6652
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6649..6665
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6649..6665
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6686..6704
                     /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 ctttgggagg
      901 gggctgtccc cgtgagctcc caggcgcgcc tgtcattcta aatctctctt tcagcctaaa
      961 gctttttccc cgtatccccc caggtgtctg caggctcaaa gagcagcgag aagcgttcag
     1021 aggaaagcga tcccgtgcca ccttccccgt gcccgggctg tccccgcacg ctgccggctc
     1081 ggggatgcgg ggggagcgcc ggaccggagc ggagccccgg gcggctcgct gctgccccct
     1141 agcgggggag ggacgtaatt acatccctgg gggctttggg ggggggctgt ccccgtgagc
     1201 tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag actgcgtcgc
     1261 ccgggtaccc gtgtatccaa taaacccctc gaggtcgacg gtatcgataa gcttgatatc
     1321 gaattcctgc agcccggggg atcacgcggc catcaagctt aggcctccac ggtggcttct
     1381 aatccgtgag tcctagcggg tgacagccct ccgtcttcac aggcggagga gagtcttccg
     1441 tagggttcct cggagtactg tcctccgacg cgtgcaaggg tcgactctag agggtatata
     1501 atggatccca tcgcgtctca gcctcacttt gagctcctcc acacgaattc cctcgacctc
     1561 gaagacgcgt ggatccaccg gtggatccct cgagcccggg tctagaggag ctactaactt
     1621 cagcctgctg aagcaggctg gagacgtgga ggagaaccct ggacctagat ccatggtgag
     1681 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1741 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1801 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1861 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     1921 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1981 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     2041 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     2101 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     2161 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     2221 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     2281 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     2341 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aatcgattct
     2401 agctagaact atagtgagtc gtattacgta gatccagaca tgataagata cattgatgag
     2461 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     2521 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     2581 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaatt cgcggccgcg
     2641 gcgactagtg ggtttattgg atacacgggt acccgggcga cgcagtctat cggaggactg
     2701 gcgcgccgag tgaggggttg tgggctcttt tattgagctc acggggacag ccccccccca
     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 cgcctgggag ctcacgggga cagccccctc ccaaagcccc cagggatgta
     3061 attacgtccc tcccccgcta gggggcagca gcgagccgcc cggggctccg ctccggtccg
     3121 gcgctccccc cgcatccccg agccggcagc gtgcggggac agcccgggca cggggaaggt
     3181 ggcacgggat cgctttcctc tgaacgcttc tcgctgctct ttgagcctgc agacacctgg
     3241 ggggatacgg ggaaaaagct ttaggctgaa agagagattt agaatgacag gcgcgccaac
     3301 tcgaattcga tatcaagctt atcgctagag gatctataac ttcgtaagat ctgtctctgc
     3361 tcacgtttcc tgctatttgc agcctctcta tcaagactaa tacacctctt cccgcatcgg
     3421 ctgcctgtga gaggcttttc agcactgcag gattgctttt cagccccaaa agagctaggc
     3481 ttgacactaa caattttgag aatcagcttc tactgaagtt aaatctgagg ttttacaact
     3541 ttgagtagcg tgtactggca ttagattgtc tgtcttatag tttgataatt aaatacaaac
     3601 agttctaaag caggataaaa ccttgtatgc atttcattta atgttttttg agattaaaag
     3661 cttaaacaag aatctctagt tttctttctt gcttttactt ttacttcctt aatactcaag
     3721 tacaatttta atggagtact tttttacttt tactcaagta agattctagc cagatacttt
     3781 tacttttaat tgagtaaaat tttccctaag tacttgtact ttcacttgag taaaattttt
     3841 gagtactttt tacacctctg gagctccaat tcgccctata gtgagtcgta ttacaattca
     3901 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     3961 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     4021 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     4081 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4141 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4201 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4261 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4321 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4381 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4441 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4501 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4561 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4621 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4681 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4741 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4801 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4861 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4921 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4981 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5041 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5101 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5161 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5221 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5281 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5341 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5401 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5461 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5521 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5581 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5641 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5701 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5761 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5821 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5881 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5941 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6001 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6061 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6121 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6181 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6241 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6301 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6361 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     6421 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     6481 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     6541 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     6601 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     6661 atgaccatga ttacgccaag ctcgaaatta accctcacta aagggaacaa aagctggtac
     6721 c
//
