LOCUS Exported 9184 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Retroviral expression vector containing Rab11a tagged with RFP. ACCESSION . VERSION . KEYWORDS pQCXIH-tag RFPt- Rab11a WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9184) AUTHORS Roman-Fernandez A, Roignot J, Sandilands E, Nacke M, Mansour MA, McGarry L, Shanks E, Mostov KE, Bryant DM TITLE The phospholipid PI(3,4)P2 is an apical identity determinant. JOURNAL Nat Commun. 2018 Nov 28;9(1):5041. doi: 10.1038/s41467-018-07464-8. PUBMED 30487552 REFERENCE 2 (bases 1 to 9184) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9184 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(39..899) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 662..681 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(900..1004) /gene="bla" /label=AmpR promoter primer_bind 1072..1090 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" enhancer 1190..1569 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1570..1773 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" LTR 1774..1949 /label=5' LTR (truncated) /note="truncated long terminal repeat from Moloney murine sarcoma virus" misc_feature 2012..2211 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 2412..2828 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 2758..2780 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 2796..2812 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" enhancer 2865..3168 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3169..3372 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 3369..3393 /label=LNCX /note="Human CMV promoter, forward primer" regulatory 3518..3527 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3524..4255 /codon_start=1 /product="photostable variant of TagRFP (Shaner et al., 2008)" /label=TagRFP-T /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV ARYCDLPSKLGHKLNGMDELYK" misc_feature 4973..5546 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(5140..5157) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5367..5386 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5585..6604 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE" LTR 7151..7576 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from Moloney murine leukemia virus" primer_bind complement(7679..7701) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(7837..7857) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 7842..8171 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 8022..8157 /label=SV40 ori /note="SV40 origin of replication" primer_bind 8084..8103 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 8193..8212 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(8293..8310) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8464..9052) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8544..8563) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 61 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 121 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 181 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 241 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 301 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc 361 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 421 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 481 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 541 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 601 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg 661 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 721 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 781 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 841 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 901 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 961 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 1021 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt 1081 atcacgaggc cctttcgtct tcaagaatta gcttggccat tgcatacgtt gtatccatat 1141 cataatatgt acatttatat tggctcatgt ccaacattac cgccatgttg acattgatta 1201 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 1261 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 1321 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 1381 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 1441 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 1501 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 1561 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 1621 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 1681 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 1741 cgtgtacggt gggaggtcta tataagcaga gctcaataaa agagcccaca acccctcact 1801 cggcgcgcca gtcttccgat agactgcgtc gcccgggtac ccgtattccc aataaagcct 1861 cttgctgttt gcatccgaat cgtggtctcg ctgttccttg ggagggtctc ctctgagtga 1921 ttgactaccc acgacggggg tctttcattt gggggctcgt ccgggatttg gagacccctg 1981 cccagggacc accgacccac caccgggagg taagctggcc agcaacttat ctgtgtctgt 2041 ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg tctgtactag ttagctaact 2101 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc 2161 ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt 2221 cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac 2281 agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc 2341 tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt 2401 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag 2461 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct 2521 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc 2581 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag 2641 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca 2701 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc 2761 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc 2821 taggcgccgg aattgaagat ctgggggatc gatcctctag agtccgttac ataacttacg 2881 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg 2941 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta 3001 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt 3061 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac 3121 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt 3181 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac 3241 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt 3301 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat 3361 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt 3421 gacctccata gaagacaccg ggaccgatcc agcctccgcg gccgggaacg gtgcattgga 3481 acggacctgc aggaattgat ccgcggccgc accggtcgcc accatggtgt ctaagggcga 3541 agagctgatt aaggagaaca tgcacatgaa gctctacatg gagggcaccg tgaacaacca 3601 ccacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag 3661 aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctaccagctt 3721 catgtacggc agcagaacct tcatcaacca cacccagggc atccccgatt tctttaagca 3781 gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct 3841 gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag 3901 aggggtgaac ttcccatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc 3961 caacaccgag atgctgtacc ccgctgacgg cggcctggaa ggcagaaccg acatggccct 4021 gaagctcgtg ggcgggggcc acctgatctg caacttcaag accacataca gatccaagaa 4081 acccgctaag aacctcaaga tgcccggcgt ctactatgtg gaccacagac tggaaagaat 4141 caaggaggcc gacaaagaga cctacgtcga gcagcacgag gtggctgtgg ccagatactg 4201 cgacctccct agcaaactgg ggcacaaact taatggcatg gacgagctgt acaagtccgg 4261 actcagatct cgagctcaag cttcgaattc aatgggcacc cgcgacgacg agtacgacta 4321 tctcttcaaa gttgtcctta ttggagattc tggtgttgga aagagtaatc tcttgtctcg 4381 atttactcga aatgagttta atctcgaaag taagagcacc attggagtag agtttgcaac 4441 aagaagcatc caggttgatg ggaaaacaat aaaggcacag atatgggaca cggcagggca 4501 ggagcgatac cgagctataa catcagcata ttatcgtgga gctgtaggtg ccttactggt 4561 ttatgacatt gctaagcatc tcacatatga aaatgtagaa cgatggctga aagaactgag 4621 agatcatgct gatagtaaca ttgttatcat gcttgtgggc aataagagtg atttgcgtca 4681 tctcagggca gttcctacag atgaagcaag agcttttgca gaaaagaatg gtttgtcatt 4741 cattgagact tccgctctag actctacaaa tgtagaagct gcttttcaga caattctgac 4801 agagatatac cgcattgttt cccagaaaca aatgtcagac agacgtgaaa atgacatgtc 4861 tccaagcaac aatgtggttc ctattcatgt tccaccaacc actgaaaaca agccaaaggt 4921 gcagtgctgt cagaacatat aaggatccgg aattccgccc cccccccccc cccccctctc 4981 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg 5041 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg 5101 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag 5161 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt 5221 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc 5281 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga 5341 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga 5401 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct 5461 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg 5521 ttttcctttg aaaaacacga tgataagctt gccacaaccc gtaccaaaga tggatagatc 5581 cggaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag 5641 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt 5701 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg 5761 ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg 5821 ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca 5881 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc 5941 gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat 6001 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca 6061 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct 6121 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc 6181 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat 6241 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg 6301 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg 6361 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg 6421 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc 6481 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg 6541 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga 6601 atagagtaga tgccgaccga acaagagctg atttcgagaa cgcctcagcc agcaactcgc 6661 gcgagcctag caaggcaaat gcgagagaac ggccttacgc ttggtggcac agttctcgtc 6721 cacagttcgc taagctcgct cggctgggtc gcgggagggc cggtcgcagt gattcaggcc 6781 cttctggatt gtgttggtcc ccagggcacg attgtcatgc ccacgcactc gggtgatctg 6841 actgatcccg cagattggag atcgccgccc gtgcctgccg attgggtgca gatctctatc 6901 gagggctgca gcgctgcaga ggccgagtgc agaactgctc caaagggacc tcaaggcttt 6961 ccgagggaca ctaggctgac tccatcgagc cagtgtagag ataagcttat cgattagtcc 7021 aatttgttaa agacaggata tcagtggtcc aggctctagt tttgactcaa caatatcacc 7081 agctgaagcc tatagagtac gagccataga taaaataaaa gattttattt agtctccaga 7141 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat 7201 tttgcaaggc atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa 7261 cagatggaac agggtcgacc ctagagaacc atcagatgtt tccagggtgc cccaaggacc 7321 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc 7381 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc 7441 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat 7501 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca 7561 gcgggggtct ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc 7621 gacccaccac cgggaggtaa gctggctgcc tcgcgcgttt cggtgatgac ggtgaaaacc 7681 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca 7741 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc 7801 agtcacgtag cgatagcgga gtgtagatcc ggctgtggaa tgtgtgtcag ttagggtgtg 7861 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 7921 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 7981 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc 8041 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg 8101 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag 8161 gcttttgcaa aaagcttact ggcttaacta tgcggcatca gagcagattg tactgagagt 8221 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 8281 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 8341 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 8401 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 8461 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 8521 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 8581 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 8641 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 8701 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 8761 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 8821 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 8881 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 8941 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 9001 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 9061 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 9121 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 9181 taaa //