LOCUS       sequence-389344-        7774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector containing GFP tagged Podocalyxin. Dual 
            mutant for HRS>AAA / lacking DTHL..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7774)
  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 7774)
  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*/PBM*
FEATURES             Location/Qualifiers
     source          1..7774
                     /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(3361..3379)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3362..3379)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3399..3416)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3405..3629
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3675..4103
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3762..3781)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3972..3993
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4112..4132)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4117..4446
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4297..4432
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4359..4378
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4513..5307
                     /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(4567..4586)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5177..5196
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5481..5602
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5518..5537)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5572..5591
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5651..5667)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5651..5667)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5664..5686)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5675..5691
                     /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(5699..5705,5706..5723,5724..5729))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5744..5765
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5882..5899)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6053..6641)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6133..6152)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6812..7603,7604..7672))
                     /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     7435..7454
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7673..7774,1..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 gagtaagcgg ccgctcgagc atgcatctag agggccctat
     3361 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     3421 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3481 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3541 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3601 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     3661 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3721 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3781 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3841 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3901 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3961 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4021 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4081 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     4141 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4201 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4261 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4321 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4381 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4441 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     4501 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4561 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4621 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4681 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4741 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4801 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4861 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4921 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4981 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5041 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5101 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5161 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5221 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5281 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5341 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5401 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5461 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     5521 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5581 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     5641 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5701 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5761 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5821 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5881 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5941 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6001 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6061 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6121 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6181 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6241 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6301 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6361 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6421 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6481 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6541 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6601 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6661 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6721 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6781 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6841 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6901 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6961 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7021 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7081 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7141 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7201 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7261 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7321 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7381 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7441 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7501 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7561 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7621 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7681 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7741 aatgtattta gaaaaataaa caaatagggg ttcc
//
