LOCUS       Exported                6053 bp ds-DNA     circular SYN 05-JUN-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6053)
  AUTHORS   Hammond Lab
  TITLE     Direct Submission
  JOURNAL   Exported Friday, May 19, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6053
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     promoter        1..539
                     /label=CMV
                     /note="CMV"
     misc_feature    1
                     /label=GFP tag
                     /note="GFP tag"
     misc_feature    1
                     /label=Kanamycin
                     /note="Kanamycin"
     promoter        519..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(519..539)
                     /label=CMV-R
     primer_bind     519..539
                     /label=CMV-F
     misc_feature    591..614
                     /label=MCS
                     /note="MCS"
     CDS             618..650
                     /codon_start=1
                     /label=Lyn11
                     /note="Lyn11"
                     /translation="MGSIKSKGKDS"
     CDS             651..665
                     /codon_start=1
                     /label=GGSGG
                     /translation="GGSGG"
     misc_feature    666..695
                     /label=Helical linker
     CDS             696..704
                     /codon_start=1
                     /label=GGSGG
                     /translation="GGS"
     CDS             705..719
                     /codon_start=1
                     /label=GGSGG
                     /translation="GGSGG"
     primer_bind     complement(715..731)
                     /label=Lyn-R
     misc_feature    720..721
                     /label=MCS
                     /note="MCS"
     primer_bind     729..746
                     /label=FPi-F
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     misc_feature    732..1445
                     /label=EGFP
                     /note="EGFP"
     primer_bind     complement(732..749)
                     /label=FPv-R
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     primer_bind     732..749
                     /label=FPi-F2.0
     primer_bind     1377..1397
                     /label=A41
     CDS             1426..1445
                     /codon_start=1
                     /label=mCherry
                     /translation="MDELYK"
     primer_bind     complement(1426..1445)
                     /label=FPi-R
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     primer_bind     1426..1445
                     /label=FPv-F
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     misc_feature    1455..1485
                     /label=MCS
                     /note="MCS"
     primer_bind     1485..1514
                     /label=PIP4K2A_SalICut_F
     CDS             1491..2711
                     /codon_start=1
                     /gene="PIP4K2A"
                     /gene_synonym="PI5P4KA; PIP5K2A; PIP5KII-alpha; PIP5KIIA;
                     PIPK"
                     /product="phosphatidylinositol 5-phosphate 4-kinase type-2
                     alpha isoform 1"
                     /EC_number="2.7.1.149"
                     /label=PIP4K2A
                     /note="isoform 1 is encoded by transcript variant 1; 
                     phosphatidylinositol-4-phosphate 5-kinase, type II, alpha; 
                     PIP5KIIalpha; PtdIns(4)P-5-kinase B isoform; type II 
                     phosphatidylinositol-4-phosphate 5-kinase 53 K isoform; 
                     1-phosphatidylinositol-4-phosphate kinase; 
                     1-phosphatidylinositol-4-phosphate-5-kinase; PIP5KIII; 
                     PIP4KII-alpha; PI(5)P 4-kinase type II alpha; 
                     ptdIns(4)P-5-kinase C isoform; diphosphoinositide kinase 
                     2-alpha; ptdIns(5)P-4-kinase isoform 2-alpha; 
                     1-phosphatidylinositol 5-phosphate 4-kinase 2-alpha; 
                     phosphatidylinositol 5-phosphate 4-kinase type II alpha"
                     /db_xref="CCDS:CCDS7141.1"
                     /db_xref="GeneID:5305"
                     /db_xref="HGNC:HGNC:8997"
                     /db_xref="MIM:603140"
                     /protein_id="NP_005019.2"
                     /translation="MATPGNLGSSVLASKTKTKKKHFVAQKVKLFRASDPLLSVLMWGV
                     NHSINELSHVQIPVMLMPDDFKAYSKIKVDNHLFNKENMPSHFKFKEYCPMVFRNLRER
                     FGIDDQDFQNSLTRSAPLPNDSQARSGARFHTSYDKRYIIKTITSEDVAEMHNILKKYH
                     QYIVECHGITLLPQFLGMYRLNVDGVEIYVIVTRNVFSHRLSVYRKYDLKGSTVAREAS
                     DKEKAKELPTLKDNDFINEGQKIYIDDNNKKVFLEKLKKDVEFLAQLKLMDYSLLVGIH
                     DVERAEQEEVECEENDGEEEGESDGTHPVGTPPDSPGNTLNSSPPLAPGEFDPNIDVYG
                     IKCHENSPRKEVYFMAIIDILTHYDAKKKAAHAAKTVKHGAGAEISTVNPEQYSKRFLD
                     FIGHILT"
     primer_bind     1491..1508
                     /label=PIP4KA_Insert_F
     CDS             2121..2123
                     /codon_start=1
                     /label=D211
                     /translation="D"
     primer_bind     2124..2148
                     /label=PIP4K2A_D211_F
     primer_bind     complement(2288..2306)
                     /label=PIP4K2A_D273_R
     CDS             2307..2309
                     /codon_start=1
                     /label=D273
                     /translation="D"
     primer_bind     2310..2334
                     /label=PIP4K2A_D273K_F
     CDS             2343..2501
                     /codon_start=1
                     /label=InsertLoop
                     /translation="RAEQEEVECEENDGEEEGESDGTHPVGTPPDSPGNTLNSSPPLAP
                     GEFDPNID"
     CDS             2574..2648
                     /codon_start=1
                     /label=Specificity Loop
                     /translation="THYDAKKKAAHAAKTVKHGAGAEIS"
     primer_bind     complement(2685..2717)
                     /label=PIP4K2A_BamHICut_R
     primer_bind     complement(2694..2711)
                     /label=PIP4KA_Insert_R
     misc_feature    2713..2748
                     /label=MCS
                     /note="MCS"
     primer_bind     complement(2747..2766)
                     /label=SV40-pArev
     misc_feature    2969..3424
                     /label=f1
                     /note="f1"
     promoter        3486..3514
                     /label=Bacterial
                     /note="Bacterial"
     promoter        3598..3827
                     /label=SV40early
                     /note="SV40early"
     misc_feature    3765..3900
                     /label=SV40
                     /note="SV40"
     misc_feature    3949..4743
                     /label=Kan/Neo
                     /note="Kan/Neo"
     misc_feature    4979..4997
                     /label=HSV-TK
                     /note="HSV-TK"
     misc_feature    5328..5971
                     /label=pUC
                     /note="pUC"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctaccatg ggatctataa aatcaaaagg gaaagacagc ggtggctctg
      661 gaggggaagc cgctgctaga gaagctgccg caagaggcgg atcagggggt tccggaggcc
      721 tcgccgccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      781 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      841 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      901 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
      961 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1021 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1081 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1141 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     1201 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     1261 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1321 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     1381 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1441 acaagtccgg actcagatct cgagctcaag cttcgaattc tgcagtcgac atggcgaccc
     1501 ccggcaacct agggtcctct gtcctggcga gcaagaccaa gaccaagaag aagcacttcg
     1561 tagcgcagaa agtgaagctg tttcgggcca gcgacccgct gctcagcgtc ctcatgtggg
     1621 gggtaaacca ctcgatcaat gaactgagcc atgttcaaat ccctgttatg ttgatgccag
     1681 atgacttcaa agcctattca aaaataaagg tggacaatca cctttttaac aaagaaaaca
     1741 tgccgagcca tttcaagttt aaggaatact gcccgatggt cttccgtaac ctgcgggaga
     1801 ggtttggaat tgatgatcaa gatttccaga attccctgac caggagcgca cccctcccca
     1861 acgactccca ggcccgcagt ggagctcgtt ttcacacttc ctacgacaaa agatacatca
     1921 tcaagactat taccagtgaa gacgtggccg aaatgcacaa catcctgaag aaataccacc
     1981 agtacatagt ggaatgtcat gggatcaccc ttcttcccca gttcttgggc atgtaccggc
     2041 ttaatgttga tggagttgaa atatatgtga tagttacaag aaatgtattc agccaccgtt
     2101 tgtctgtgta taggaaatac gacttaaagg gctctacagt ggctagagaa gctagtgaca
     2161 aagaaaaggc caaagaactg ccaactctga aagataatga tttcattaat gagggccaaa
     2221 agatttatat tgatgacaac aacaagaagg tcttcctgga aaaactaaaa aaggatgttg
     2281 agtttctggc ccagctgaag ctcatggact acagtctgct ggtgggaatt catgatgtgg
     2341 agagagccga acaggaggaa gtggagtgtg aggagaacga tggggaggag gagggcgaga
     2401 gcgatggcac ccacccggtg ggaacccccc cagatagccc cgggaataca ctgaacagct
     2461 caccacccct ggctcccggg gagttcgatc cgaacatcga cgtctatgga attaagtgcc
     2521 atgaaaactc gcctaggaag gaggtgtact tcatggcaat tattgacatc cttactcatt
     2581 atgatgcaaa aaagaaagct gcccatgctg caaaaactgt taaacatggc gctggcgcgg
     2641 agatctccac cgtgaaccca gaacagtatt caaagcgctt tttggacttt attggccaca
     2701 tcttgacgta aggatccacc ggatctagat aactgatcat aatcagccat accacatttg
     2761 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     2821 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     2881 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     2941 ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc
     3001 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     3061 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     3121 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     3181 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     3241 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     3301 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     3361 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     3421 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3481 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3541 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     3601 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3661 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3721 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3781 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3841 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3901 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3961 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4021 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4081 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     4141 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4201 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4261 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4321 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4381 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4441 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     4501 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4561 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4621 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4681 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4741 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4801 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4861 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     4921 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     4981 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     5041 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     5101 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     5161 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     5221 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5281 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5341 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5401 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5461 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     5521 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5581 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5641 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5701 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5761 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5821 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5881 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5941 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6001 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
