LOCUS       Exported                8278 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression vector, containing a GFP tagged version of 
            INPP5D.
ACCESSION   .
VERSION     .
KEYWORDS    INPP5D-eGFP WT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8278)
  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 8278)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8278
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        140..443
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        444..647
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     598..618
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      676..685
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     RBS             2275..2283
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     regulatory      4297..4306
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4303..5022
                     /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             4303..5019
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4348..4369)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4609..4628)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4956..4977
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5145..5266
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5182..5201)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5236..5255
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5273..5728)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5410..5431)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5622..5641
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5755..5859
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5861..6218
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(5884..5904)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6069..6204
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6131..6150
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6253..7047
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6307..6326)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6917..6936
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7235..7254)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7279..7326
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7655..8243
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8144..8163
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
       61 ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata
      121 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      181 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      241 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      301 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      361 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      421 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      481 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      541 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      601 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
      661 gtcagatccg ctagcgccac catggtcccc tgctggaacc atggcaacat cacccgctcc
      721 aaggcggagg agctgctttc caggacaggc aaggacggga gcttcctcgt gcgtgccagc
      781 gagtccatct cccgggcata cgcgctctgc gtgctgtatc ggaattgcgt ttacacttac
      841 agaattctgc ccaatgaaga tgataaattc actgttcagg catccgaagg cgtctccatg
      901 aggttcttca ccaagctgga ccagctcatc gagttttaca agaaggaaaa catggggctg
      961 gtgacccatc tgcaataccc tgtgccgctg gaggaagagg acacaggcga cgaccctgag
     1021 gaggacacag aaagtgtcgt gtctccaccc gagctgcccc caagaaacat cccgctgact
     1081 gccagctcct gtgaggccaa ggaggttcct ttttcaaacg agaatccccg agcgaccgag
     1141 accagccggc cgagcctctc cgagacattg ttccagcgac tgcaaagcat ggacaccagt
     1201 gggcttccag aagagcatct taaggccatc caagattatt taagcactca gctcgcccag
     1261 gactctgaat ttgtgaagac agggtccagc agtcttcctc acctgaagaa actgaccaca
     1321 ctgctctgca aggagctcta tggagaagtc atccggaccc tcccatccct ggagtctctg
     1381 cagaggttat ttgaccagca gctctccccg ggcctccgtc cacgtcctca ggttcctggt
     1441 gaggccaatc ccatcaacat ggtgtccaag ctcagccaac tgacaagcct gttgtcgtcc
     1501 attgaagaca aggtcaaggc cttgctgcac gagggtcctg agtctccgca ccggccctcc
     1561 cttatccctc cagtcacctt tgaggtgaag gcagagtctc tggggattcc tcagaaaatg
     1621 cagctcaaag tcgacgttga gtctgggaaa ctgatcatta agaagtccaa ggatggttct
     1681 gaggacaagt tctacagcca caagaaaatc ctgcagctga ttaagtcaca gaaatttctg
     1741 aataagttgg tgatcttggt ggaaacagag aaggagaaga tcctgcggaa ggaatatgtt
     1801 tttgctgact ccaaaaagag agaaggcttc tgccagctcc tgcagcagat gaagaacaag
     1861 cactcagagc agccggagcc cgacatgatc accatcttca tcggcacctg gaacatgggt
     1921 aacgcccccc ctcccaagaa gatcacgtcc tggtttctct ccaaggggca gggaaagacg
     1981 cgggacgact ctgcggacta catcccccat gacatttacg tgatcggcac ccaagaggac
     2041 cccctgagtg agaaggagtg gctggagatc ctcaaacact ccctgcaaga aatcaccagt
     2101 gtgactttta aaacagtcgc catccacacg ctctggaaca tccgcatcgt ggtgctggcc
     2161 aagcctgagc acgagaaccg gatcagccac atctgtactg acaacgtgaa gacaggcatt
     2221 gcaaacacac tggggaacaa gggagccgtg ggggtgtcgt tcatgttcaa tggaacctcc
     2281 ttagggttcg tcaacagcca cttgacttca ggaagtgaaa agaaactcag gcgaaaccaa
     2341 aactatatga acattctccg gttcctggcc ctgggcgaca agaagctgag tccctttaac
     2401 atcactcacc gcttcacgca cctcttctgg tttggggatc ttaactaccg tgtggatctg
     2461 cctacctggg aggcagaaac catcatccag aaaatcaagc agcagcagta cgcagacctc
     2521 ctgtcccacg accagctgct cacagagagg agggagcaga aggtcttcct acacttcgag
     2581 gaggaagaaa tcacgtttgc cccaacctac cgttttgaga gactgactcg ggacaaatac
     2641 gcctacacca agcagaaagc gacagggatg aagtacaact tgccttcctg gtgtgaccga
     2701 gtcctctgga agtcttatcc cctggtgcac gtggtgtgtc agtcttatgg cagtaccagc
     2761 gacatcatga cgagtgacca cagccctgtc tttgccacat ttgaggcagg agtcacttcc
     2821 cagtttgtct ccaagaacgg tcccgggact gttgacagcc aaggacagat tgagtttctc
     2881 aggtgctatg ccacattgaa gaccaagtcc cagaccaaat tctacctgga gttccactcg
     2941 agctgcttgg agagttttgt caagagtcag gaaggagaaa atgaagaagg aagtgagggg
     3001 gagctggtgg tgaagtttgg tgagactctt ccaaagctga agcccattat ctctgaccct
     3061 gagtacctgc tagaccagca catcctcatc agcatcaagt cctctgacag cgacgaatcc
     3121 tatggcgagg gctgcattgc ccttcggtta gaggccacag aaacgcagct gcccatctac
     3181 acgcctctca cccaccatgg ggagttgaca ggccacttcc agggggagat caagctgcag
     3241 acctctcagg gcaagacgag ggagaagctc tatgactttg tgaagacgga gcgtgatgaa
     3301 tccagtgggc caaagaccct gaagagcctc accagccacg accccatgaa gcagtgggaa
     3361 gtcactagca gggcccctcc gtgcagtggc tccagcatca ctgaaatcat caaccccaac
     3421 tacatgggag tggggccctt tgggccacca atgcccctgc acgtgaagca gaccttgtcc
     3481 cctgaccagc agcccacagc ctggagctac gaccagccgc ccaaggactc cccgctgggg
     3541 ccctgcaggg gagaaagtcc tccgacacct cccggccagc cgcccatatc acccaagaag
     3601 tttttaccct caacagcaaa ccggggtctc cctcccagga cacaggagtc aaggcccagt
     3661 gacctgggga agaacgcagg ggacacgctg cctcaggagg acctgccgct gacgaagccc
     3721 gagatgtttg agaaccccct gtatgggtcc ctgagttcct tccctaagcc tgctcccagg
     3781 aaggaccagg aatcccccaa aatgccgcgg aaggaacccc cgccctgccc ggaacccggc
     3841 atcttgtcgc ccagcatcgt gctcaccaaa gcccaggagg ctgatcgcgg cgaggggccc
     3901 ggcaagcagg tgcccgcgcc ccggctgcgc tccttcacgt gctcatcctc tgccgagggc
     3961 agggcggccg gcggggacaa gagccaaggg aagcccaaga ccccggtcag ctcccaggcc
     4021 ccggtgccgg ccaagaggcc catcaagcct tccagatcgg aaatcaacca gcagaccccg
     4081 cccaccccga cgccgcggcc gccgctgcca gtcaagagcc cggcggtgct gcacctccag
     4141 cactccaagg gccgcgacta ccgcgacaac accgagctcc cgcatcacgg caagcaccgg
     4201 ccggaggagg ggccaccagg gcctctaggc aggactgcca tgcagaagct tcgaattctg
     4261 cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca ccatggtgag caagggcgag
     4321 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     4381 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     4441 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     4501 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     4561 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     4621 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     4681 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     4741 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     4801 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     4861 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     4921 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     4981 gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg cgactctaga
     5041 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     5101 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     5161 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5221 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     5281 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     5341 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     5401 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     5461 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     5521 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     5581 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     5641 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     5701 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5761 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5821 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     5881 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     5941 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     6001 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     6061 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     6121 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     6181 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     6241 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     6301 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     6361 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     6421 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     6481 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     6541 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     6601 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     6661 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     6721 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     6781 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     6841 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     6901 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     6961 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     7021 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     7081 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     7141 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     7201 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     7261 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     7321 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     7381 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     7441 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     7501 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     7561 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     7621 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     7681 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     7741 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     7801 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     7861 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     7921 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7981 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     8041 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     8101 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     8161 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     8221 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggc
//
