LOCUS       mEGFP_(A206K)-PT        6612 bp DNA     circular SYN 24-SEP-2024
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6612)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 24, 2024 from SnapGene 7.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    593..612
                     /label=Feature 18
                     /note="color: #a6acb3"
     CDS             613..1329
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1:  613 ..  615 / #05fd14
                        2:  616 ..  618 / #05fd14 / 1a
                        3:  619 .. 1329 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
                     /note="color: black; sequence: CGTCGCCGTCCAGCTCGACCAG"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
                     /note="color: black; sequence: GTCTTGTAGTTGCCGTCGTC"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
                     /note="color: black; sequence: CATGGTCCTGCTGGAGTTCGTG"
     misc_feature    1351..2559
                     /label=PTEN
                     /note="color: #a6acb3"
     primer_bind     complement(1359..1379)
                     /label=R14G REV
                     /note="color: black; sequence: ctaacgatctctttgatgatg; 
                     added: 2022-01-11"
     primer_bind     1380..1402
                     /label=R14G FOR
                     /note="color: black; sequence: cagaaacaaaGggagatatcaag; 
                     added: 2022-01-11"
     CDS             1390..1392
                     /codon_start=1
                     /label=R14G A-->G (LOF)
                     /note="color: #993366"
                     /translation="G"
     primer_bind     2500..2519
                     /label=PTEN_C_For
                     /note="color: black; sequence: gactctgatccagagaatga; added:
                     2021-07-15"
     misc_feature    2538..2559
                     /label=OVERLAP
                     /note="color: #a6acb3"
     misc_feature    2560..2568
                     /label=linker PTP
                     /note="color: #a6acb3"
     misc_feature    2569..3282
                     /label=sREACh
                     /note="color: #a6acb3"
     primer_bind     complement(2611..2632)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
                     /note="color: black; sequence: CGTCGCCGTCCAGCTCGACCAG"
     primer_bind     complement(2872..2891)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
                     /note="color: black; sequence: GTCTTGTAGTTGCCGTCGTC"
     primer_bind     3219..3234
                     /label=EGFP-C
                     /note="EGFP, forward primer"
                     /note="color: black; sequence: CATGGTCCTGCTGGAGTTCGTG"
     primer_bind     3286..3306
                     /label=V_FOR_ALT
                     /note="color: black; sequence: AGATAACTGATCATAATCAGC; 
                     added: 2021-11-16"
     polyA_signal    3400..3521
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      complement(3528..3983)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        4010..4114
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     promoter        4116..4473
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      4324..4459
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             4508..5302
                     /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)"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    5534..5581
                     /label=HSV TK poly(A) signal
                     /note="herpesvirus thymidine kinase polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      5910..6498
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
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 aagctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct atgacagcca tcatcaaaga gatcgttagc
     1381 agaaacaaag ggagatatca agaggatgga ttcgacttag acttgaccta tatttatcca
     1441 aatattattg ctatgggatt tcctgcagaa agacttgaag gtgtatacag gaacaatatt
     1501 gatgatgtag taaggttttt ggattcaaag cataaaaacc attacaagat atacaatcta
     1561 tgtgctgaga gacattatga caccgccaaa tttaactgca gagttgcaca gtatcctttt
     1621 gaagaccata acccaccaca gctagaactt atcaaaccct tttgtgaaga tcttgaccaa
     1681 tggctaagtg aagacgacaa tcatgttgca gcaattcact gtaaagctgg gaaaggacgg
     1741 actggtgtaa tgatttgtgc atatttattg catcggggca agtttttaaa ggcacaagag
     1801 gccctggatt tttatgggga agtaaggacc agagataaaa agggagtaac tattcccagt
     1861 cagaggcgct atgtatatta ttatagctac ctgttaaaga atcacctgga ttacagacca
     1921 gtggcactgt tgtttcacaa gatgatgttt gaaactattc caatgttcag tggcggaact
     1981 tgcaatcccc agtttgtggt ctgccagcta aaggtgaaga tctactcctc caactcagga
     2041 cccacgcggc gggaggacaa gctcatgtac tttgagttcc ctcagccatt gcctgtgtgt
     2101 ggtgacatca aagtagagtt cttccacaaa cagaacaaga tgctcaaaaa ggacaaaatg
     2161 tttcactttt gggtaaatac gttcttcata ccaggaccag aggaaacctc agaaaaagtg
     2221 gaaaatggaa gtctttgtga tcaggaaatc gatagcattt gtagtataga gcgtgcggat
     2281 aatgacaagg agtatcttgt gctcaccctg acaaaaaatg atcttgacaa agcaaacaaa
     2341 gacaaggcca accgatactt ctctccaaat tttaaggtga aattatactt tacaaaaaca
     2401 gtagaggaac catcaaatcc agaggctagc agttcaactt ctgtgactcc agacgttagt
     2461 gacaatgaac ctgatcatta tagatattct gacaccactg actctgatcc agagaatgaa
     2521 ccttttgatg aagatcagca ttcacaaatt acaaaagtcc ctactcctgt gagcaagggc
     2581 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2641 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     2701 aagctcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc
     2761 ggctacggcc tgatgtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     2821 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     2881 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2941 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     3001 tggaacagcg tcaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     3061 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     3121 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     3181 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagagagtg
     3241 accgccgccg ggatcactct cggcatggac gagctgtaca agtgaagata actgatcata
     3301 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     3361 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     3421 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3481 cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt
     3541 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     3601 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     3661 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3721 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     3781 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     3841 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     3901 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3961 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4021 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4081 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     4141 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4201 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4261 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4321 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4381 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4441 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     4501 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4561 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4621 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4681 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4741 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4801 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4861 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4921 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4981 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     5041 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5101 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5161 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5221 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5281 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     5341 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5401 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5461 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     5521 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     5581 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     5641 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     5701 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     5761 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5821 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5881 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5941 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6001 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     6061 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6121 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6181 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6241 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6301 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6361 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6421 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6481 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6541 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6601 accgccatgc at
//
