LOCUS       Exported                6396 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  This vector is used for cloning a gene driven by 4xnrUAS, to be 
            co-expressed with mRFP through P2A self-cleaving peptides..
ACCESSION   .
VERSION     .
KEYWORDS    pBT2-4xUAS:P2A-mRFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6396)
  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 6396)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6396
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     polyA_signal    1089..1223
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1126..1145)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1180..1199
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1228..1245
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(1262..1939)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1324..1347)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1522..1541)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1791..1809
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(1946..2002)
                     /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"
     primer_bind     complement(2306..2322)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2307..2323)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3549..3568)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3550..3568)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3575..3592)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3575..3591)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3584..3606)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3733..4188
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3820..3839)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4030..4051
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4214..4318
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4319..5179
                     /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(4537..4556)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5350..5938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5839..5858
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6092..6109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6226..6247
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6262..6292
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6300..6316
                     /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     6305..6327
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6324..6340
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6324..6340
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6361..6379
                     /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 acccactagt cgccgcggcc
     1021 gcgaattaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     1081 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     1141 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     1201 atgtatctta tcatgtctgg atctacgtaa tacgactcac tatagttcta gctagaatcg
     1261 attaggcgcc ggtggagtgg cggccctcgg cgcgctcgta ctgttccacg atggtgtagt
     1321 cctcgttgtg ggaggtgatg tccagcttga tgtcggtctt gtaggcgccg ggcagctgca
     1381 cgggcttctt ggccatgtag gtggtcttga cctcggcgtc gtagtggccg ccgtccttca
     1441 gcttcagcct catcttgatc tcgcccttca gggcgccgtc ctcggggtac atccgctcgg
     1501 tggaggcctc ccagcccatg gtcttcttct gcattacggg gccgtcggag gggaagttgg
     1561 tgccgcgcag cttcaccttg tagatgaact cgccgtcctg cagggaggag tcctgggtca
     1621 cggtcaccac gccgccgtcc tcgaagttca tcacgcgctc ccacttgaag ccctcgggga
     1681 aggacagctt caagtagtcg gggatgtcgg cggggtgctt cacgtaggcc ttggagccgt
     1741 actggaactg aggggacagg atgtcccagg cgaagggcag ggggccgccc ttggtcacct
     1801 tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc gccctcgccc tcgatctcga
     1861 actcgtggcc gttcacggag ccctccatgc gcaccttgaa gcgcatgaac tccttgatga
     1921 cgtcctcgga ggaggccatg gatctaggtc cagggttctc ctccacgtct ccagcctgct
     1981 tcagcaggct gaagttagta gctcctctag acccgggctc gagggatcca ccggtggatc
     2041 cacgcgtctt cgaggtcgag ggaattcgtg tggaggagct caaagtgagg ctgagacgcg
     2101 atgggatcca ttatataccc tctagagtcg acccttgcac gcgtcggagg acagtactcc
     2161 gaggaaccct acggaagact ctcctccgcc tgtgaagacg gagggctgtc acccgctagg
     2221 actcacggat tagaagccac cgtggaggcc taagcttgat ggccgcgtga tcccccgggc
     2281 tgcaggaatt cgatatcaag cttatcgata ccgtcgacct cgaggggttt attggataca
     2341 cgggtacccg ggcgacgcag tctatcggag gactggcgcg ccgagtgagg ggttgtgggc
     2401 tcttttattg agctcacggg gacagccccc ccccaaagcc cccagggatg taattacgtc
     2461 cctcccccgc tagggggcag cagcgagccg cccggggctc cgctccggtc cggcgctccc
     2521 cccgcatccc cgagccggca gcgtgcgggg acagcccggg cacggggaag gtggcacggg
     2581 atcgctttcc tctgaacgct tctcgctgct ctttgagcct gcagacacct ggggggatac
     2641 ggggaaaaag ctttaggctg aaagagagat ttagaatgac aggcgcgcct gggagctcac
     2701 ggggacagcc ccctcccaaa gcccccaggg atgtaattac gtccctcccc cgctaggggg
     2761 cagcagcgag ccgcccgggg ctccgctccg gtccggcgct ccccccgcat ccccgagccg
     2821 gcagcgtgcg gggacagccc gggcacgggg aaggtggcac gggatcgctt tcctctgaac
     2881 gcttctcgct gctctttgag cctgcagaca cctgggggga tacggggaaa aagctttagg
     2941 ctgaaagaga gatttagaat gacaggcgcg ccaactcgaa ttcgatatca agcttatcgc
     3001 tagaggatct ataacttcgt aagatctgtc tctgctcacg tttcctgcta tttgcagcct
     3061 ctctatcaag actaatacac ctcttcccgc atcggctgcc tgtgagaggc ttttcagcac
     3121 tgcaggattg cttttcagcc ccaaaagagc taggcttgac actaacaatt ttgagaatca
     3181 gcttctactg aagttaaatc tgaggtttta caactttgag tagcgtgtac tggcattaga
     3241 ttgtctgtct tatagtttga taattaaata caaacagttc taaagcagga taaaaccttg
     3301 tatgcatttc atttaatgtt ttttgagatt aaaagcttaa acaagaatct ctagttttct
     3361 ttcttgcttt tacttttact tccttaatac tcaagtacaa ttttaatgga gtactttttt
     3421 acttttactc aagtaagatt ctagccagat acttttactt ttaattgagt aaaattttcc
     3481 ctaagtactt gtactttcac ttgagtaaaa tttttgagta ctttttacac ctctggagct
     3541 ccaattcgcc ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg
     3601 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     3661 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     3721 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3781 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3841 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3901 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3961 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4021 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4081 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4141 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     4201 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4261 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4321 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4381 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4441 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4501 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4561 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4621 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4681 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4741 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4801 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4861 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4921 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4981 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5041 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5101 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5161 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     5221 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5281 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5341 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5401 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5461 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5521 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5581 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5641 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5701 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5761 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5821 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5881 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5941 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6001 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     6061 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     6121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     6181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     6241 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     6301 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagctcga
     6361 aattaaccct cactaaaggg aacaaaagct ggtacc
//
