LOCUS       Exported                6354 bp ds-DNA     circular SYN 21-OCT-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6354)
  AUTHORS   RWills
  TITLE     Direct Submission
  JOURNAL   Exported Friday, May 19, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6354
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..589
                     /label=CMV
                     /note="CMV"
     misc_feature    1
                     /label=Kanamycin
                     /note="Kanamycin"
     misc_feature    1
                     /label=GFP tag
                     /note="GFP tag"
     primer_bind     519..539
                     /label=CMV-F
     regulatory      607..616
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             613..1329
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     613..630
                     /label=FPi-F
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     primer_bind     616..633
                     /label=FPi-F2.0
     primer_bind     complement(616..633)
                     /label=FPv-R
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     primer_bind     1261..1281
                     /label=A41
     primer_bind     complement(1309..1329)
                     /label=RepGFPr
     primer_bind     1310..1329
                     /label=FPv-F
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     primer_bind     complement(1310..1329)
                     /label=FPi-R
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     misc_feature    1330..1383
                     /label=MCS
                     /note="MCS"
     primer_bind     1330..1349
                     /label=RepGFPf
     primer_bind     complement(1354..1374)
                     /label=INPPL1_V_R
     primer_bind     1357..1380
                     /label=Amp_PIP5K1B_F
     primer_bind     complement(1364..1401)
                     /label=EGFP_PIP5K1B_vR
     CDS             1384..3006
                     /codon_start=1
                     /label=HsPIP5K1B
                     /note="unnamed protein product; highly similar to Homo
                     sapiens phosphatidylinositol-4-phosphate 5-kinase, type I, 
                     beta (PIP5K1B), transcript variant 2, mRNA"
                     /protein_id="BAF85423.1"
                     /translation="MSSAAENGEAAPGKQNEEKTYKKTASSAIKGAIQLGIGYTVGNLT
                     SKPERDVLMQDFYVVESVFLPSEGSNLTPAHHYPDFRFKTYAPLAFRYFRELFGIKPDD
                     YLYSICSEPLIELSNPGASGSLFFVTSDDEFIIKTVQHKEAEFLQKLLPGYYMNLNQNP
                     RTLLPKFYGLYCMQSGGINIRIVVMNNVLPRSMRMHFTYDLKGSTYKRRASRKEREKSN
                     PTFKDLDFLQDMHEGLYFDTETYNALMKTLQRDCRVLESFKIMDYSLLLGIHFLDHSLK
                     EKEEETPQNVPDAKRTGMQKVLYSTAMESIQGPGKSGDGIITENPDTMGGIPAKSHRGE
                     KLLLFMGIIDILQSYRLMKKLEHSWKALVYDGDTVSVHRPSFYADRFLKFMNSRVFKKI
                     QALKASPSKKRCNSIAALKATSQEIVSSISQEWKDEKRDLLTEGQSFSSLDEEALGSRH
                     RPDLVPSTPSLFEAASLATTISSSSLYVNEHYPHDRPTLYSNSKGLPSSSTFTLEEGTI
                     YLTAEPNTLEVQDDNASVLDVYL"
     CDS             1384..1467
                     /codon_start=1
                     /label=NTerm
                     /translation="MSSAAENGEAAPGKQNEEKTYKKTASSA"
     primer_bind     complement(2151..2178)
                     /label=PIP5K1B_D266_R
     CDS             2179..2181
                     /codon_start=1
                     /label=D266
                     /translation="D"
     primer_bind     2182..2215
                     /label=PIP5K1B_D266K_F
     CDS             2215..2367
                     /codon_start=1
                     /label=InsertLoop
                     /translation="HSLKEKEEETPQNVPDAKRTGMQKVLYSTAMESIQGPGKSGDGII
                     TENPDT"
     CDS             2440..2511
                     /codon_start=1
                     /label=Activation Loop
                     /translation="QSYRLMKKLEHSWKALVYDGDTVS"
     CDS             2572..2577
                     /codon_start=1
                     /label=KK1B
                     /translation="KK"
     primer_bind     complement(2994..3035)
                     /label=AmpSma_PIP5K1B_R
     primer_bind     complement(2996..3035)
                     /label=Amp_PIP5K1B_R
     misc_feature    3007..3040
                     /label=MCS
                     /note="MCS"
     primer_bind     3011..3031
                     /label=EGFP_PIP5K1B_vF
     primer_bind     complement(3048..3067)
                     /label=SV40-pArev
     primer_bind     3101..3123
                     /label=A42
     misc_feature    3270..3725
                     /label=f1
                     /note="f1"
     promoter        3787..3815
                     /label=Bacterial
                     /note="Bacterial"
     promoter        3899..4128
                     /label=SV40early
                     /note="SV40early"
     misc_feature    4066..4201
                     /label=SV40
                     /note="SV40"
     misc_feature    4250..5044
                     /label=Kan/Neo
                     /note="Kan/Neo"
     misc_feature    5280..5298
                     /label=HSV-TK
                     /note="HSV-TK"
     misc_feature    5629..6272
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc
     1381 gcgatgtctt ctgctgctga aaatggagag gcagcacctg gaaaacaaaa tgaagaaaaa
     1441 acctataaaa agactgcatc atctgctatt aaaggtgcta ttcagctggg aataggatac
     1501 acagtgggta atctcacttc caagccagaa cgagatgttc ttatgcaaga cttttatgtg
     1561 gtggaaagtg tgttcctacc cagcgaaggg agcaatctga ccccagcaca tcactaccca
     1621 gactttagat ttaagacata cgctccatta gcattccgat atttcagaga actttttggt
     1681 atcaagcctg atgattactt gtattccatc tgcagtgaac ctctaataga actgtctaac
     1741 cctggagcca gtggatcctt gttttttgtg accagtgatg atgaatttat catcaaaaca
     1801 gttcagcaca aagaagctga gtttcttcag aagctactgc caggctatta catgaattta
     1861 aaccagaatc caaggactct tttgccaaaa ttttacggac tgtattgtat gcaatcagga
     1921 ggcattaata tcaggattgt ggtgatgaac aacgttttgc cacgctccat gagaatgcac
     1981 tttacatatg acttgaaagg ctcaacgtat aagcgaagag catcccgtaa agagagagag
     2041 aaatccaacc ccacatttaa ggacttagat ttcctgcaag acatgcacga agggttgtat
     2101 tttgatacgg aaacatacaa cgcgcttatg aaaacacttc agagagactg ccgggtgcta
     2161 gaaagcttca agatcatgga ttatagcctt ctgttgggaa ttcatttcct ggaccattcc
     2221 ctcaaagaga aagaggagga gaccccacaa aatgtgcctg atgctaagcg gactgggatg
     2281 cagaaggttc tctactcaac agccatggaa tctatccagg gtccagggaa atctggagat
     2341 gggataatca cagagaaccc agacacaatg ggaggcattc cagctaaaag ccatagggga
     2401 gaaaaactac ttttatttat gggcattatt gacattctgc aatcatatag gttaatgaag
     2461 aagttagaac attcctggaa agctcttgtt tatgatgggg acactgtttc tgttcataga
     2521 ccaagctttt atgcagacag atttcttaag ttcatgaatt ccagagtttt caagaaaatt
     2581 caagctttga aggcttcacc gtctaagaaa cggtgcaatt caatcgccgc cctaaaggcc
     2641 acttcacagg agattgtgtc ctcaattagc caggaatgga aggatgagaa gcgggatttg
     2701 ctgactgaag gacaaagttt tagcagcctt gatgaagaag ccctgggatc ccgacacagg
     2761 ccagacctgg tccctagcac tccatcactg tttgaagctg cttccttggc aaccacaatt
     2821 tcatcttctt ccttatacgt caatgagcac tatccacacg acaggcctac actctattca
     2881 aacagcaaag ggttaccttc cagttcaaca tttaccttgg aagaggggac catctacttg
     2941 accgctgagc ccaacactct ggaagtgcag gatgacaatg cttctgtgct tgacgtctat
     3001 ttataacgcc cgggatccac cggatctaga taactgatca taatcagcca taccacattt
     3061 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     3121 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     3181 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3241 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt
     3301 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     3361 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     3421 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     3481 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     3541 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     3601 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     3661 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     3721 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3781 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3841 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     3901 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3961 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4021 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4081 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4141 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4201 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4261 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4321 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4381 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     4441 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4501 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4561 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4621 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4681 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4741 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     4801 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4861 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4921 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4981 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     5041 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5101 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5161 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     5221 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     5281 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     5341 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     5401 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     5461 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     5521 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5581 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5641 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5701 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5761 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     5821 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5881 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5941 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6001 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6061 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6121 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6181 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6241 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6301 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
