LOCUS       sequence-384082-        7786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector containing GFP tagged Podocalyxin. With 
            H477/R479/S481>AAA mutation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7786)
  AUTHORS   Bryant DM, Roignot J, Datta A, Overeem AW, Kim M, Yu W, Peng X, 
            Eastburn DJ, Ewald AJ, Werb Z, Mostov KE
  TITLE     A molecular switch for the orientation of epithelial cell 
            polarization.
  JOURNAL   Dev Cell. 2014 Oct 27;31(2):171-87. doi: 
            10.1016/j.devcel.2014.08.027. Epub 2014 Oct 9.
  PUBMED    25307480
REFERENCE   2  (bases 1 to 7786)
  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: 
            "10.1016/j.devcel.2014.08.027"; journalName: "Dev Cell"; date: 
            "2014-10-27- 27"; volume: "31"; issue: "2"; pages: "171-87"
COMMENT     Sequence Label: GFP-PODXL FBM*
FEATURES             Location/Qualifiers
     source          1..7786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(82..101)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        273..652
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        653..856
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     807..827
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     901..920
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        901..919
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1002..1034
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     CDS             join(1035..1037,1038..1040,1041..1751)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMTDKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1080..1101)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1341..1360)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1688..1709
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(3373..3391)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3374..3391)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3411..3428)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3417..3641
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3687..4115
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3774..3793)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3984..4005
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4124..4144)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4129..4458
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4309..4444
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4371..4390
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4525..5319
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4579..4598)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5189..5208
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5493..5614
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5530..5549)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5584..5603
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5663..5679)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5663..5679)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5676..5698)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5687..5703
                     /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)."
     promoter        complement(join(5711..5717,5718..5735,5736..5741))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5756..5777
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5894..5911)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6065..6653)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6145..6164)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6824..7615,7616..7684))
                     /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     7447..7466
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7685..7786,1..3))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc gatctcccga
       61 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
      121 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
      181 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
      241 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
      301 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      361 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      421 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      481 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      541 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      601 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      661 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      721 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      781 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      841 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
      901 taatacgact cactataggg agacccaagc ttggtaccat ggccttacca gtgaccgcct
      961 tgctcctgcc gctagccttg ctgctgcatg ccgccaggcc gtatactgat atcgaaatga
     1021 atcgattggg taagatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1081 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1141 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1201 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     1261 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1321 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1381 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1441 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatgaccg
     1501 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     1561 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1621 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     1681 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     1741 agctgtacaa gaatggcaca gaaggcccta acttctacgt gcccttctcc aatgcgacgg
     1801 aattccagga aaagagtccc cagcccggcc caacccccat ggccacttca acatctacca
     1861 ggcctgcccc agcctcagcc ccagccccca aaagctccgt ggcggcttcg gtcccggccg
     1921 agcagaacac tacacccatg acgacgaaag ccccagccac ccagagcccc tctgcatccc
     1981 ctggaagctc cgtggagaac tcagccccgg cccagggctc gaccacaaca cagcagagcc
     2041 tgagtgttac tactaaggct gaagctaagg atgcaggtgg cgtccccacg gcacacgtca
     2101 ccggctctgc tcggcctgtc accagtggca gccaggtcgc agctcaggac ccagctgcat
     2161 ccaaagctcc gagcaaccac agcatcacca caaagcctct ggccacagag gccacgagcc
     2221 aggccccccg tcagaccaca gatgtcggca ccccgggacc cactgctcct cctgtgacca
     2281 actcgaccag ccctgacctg ctgggccatg ccaccccgaa gccctccgag gggccccagc
     2341 tcagtttccc cacggctgct ggcagcctgg gccctgtcac tggctctggc accggctctg
     2401 gcaccctctc cacaccccag gggaagcctg cgacactgac gcccgtggcc agctcagcag
     2461 agactcaagg gatgcccagc ccgatgccac ccagcccagc ctccccctcc tcctccccct
     2521 ttccctcctc tccctccccc tcccccgcac tgcagccctc gggcccctcg gcggcaggaa
     2581 cagaggacac aaccggcagg ggccccacaa gctccagcac tgagttggct tcaactgccc
     2641 tgcacggccc cagcacgctt tcccccacct cggcagtgag agaccagagg gtaagctgcg
     2701 ggcctccgga gagaccaacc gagcagctgc tcatcctgaa cctcaccaga agcagcccct
     2761 gcacagacag ccttccggaa gacaaactgg tcaccctgct gtgccgagcg gccaagccca
     2821 ccttcaaccc cgcccaggac cagtgccacg tgctgctggc gcccatgctg gggagccacg
     2881 cggtggtggt caaagaaatc accatcaaga cgaatctgct gcccacggcc gtgttcgagc
     2941 tgctgaagga cagatgggat gacctgagag aggagggcgt cagcgacatg cagctggggg
     3001 accaggggcc cccggaggag accgaggacc gcttcagcct gccgctcatc atcaccatcg
     3061 tctgcatggc ctccttcctg ctgcttgtcg cggccctcta cggctgctgc gcccaggccc
     3121 tcgcccatag gaaggaccag caacgactca cggaggagct gcagaccgtg gagaacggtt
     3181 accatgacaa cccgacccta gaagtgatgg agacgtcggc cgagatgcag gagaagaagg
     3241 tggtcaacct taacggggag ctgggggaca gctggattgt cccgctggac aacctgacca
     3301 aggacgacct ggacgaggag gaggacacgc acctctaagc ggccgctcga gcatgcatct
     3361 agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt
     3421 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3481 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3541 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3601 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3661 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3721 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3781 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3841 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3901 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3961 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4021 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4081 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4141 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4201 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4261 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4321 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4381 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4441 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4501 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4561 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4621 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4681 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4741 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4801 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4861 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4921 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4981 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5041 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5101 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5161 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5221 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5281 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5341 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5401 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5461 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5521 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5581 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5641 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5701 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5761 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5821 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5881 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5941 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6001 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6061 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6121 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6181 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6241 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6301 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6361 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6421 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6481 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6541 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6601 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6661 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6721 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6781 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6841 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6901 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6961 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7021 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7081 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7141 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7201 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7261 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7321 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7381 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7441 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7501 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7561 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7621 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7681 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7741 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttcc
//
