LOCUS       sequence-238532-        5539 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5539)
  AUTHORS   Mosessian S, Avliyakulov NK, Mulholland DJ, Boontheung P, Loo JA, Wu
            H
  TITLE     Analysis of PTEN complex assembly and identification of 
            heterogeneous nuclear ribonucleoprotein C as a component of the 
            PTEN-associated complex.
  JOURNAL   J Biol Chem. 2009 Oct 30;284(44):30159-66. doi: 
            10.1074/jbc.M109.027995. Epub 2009 Sep 9.
  PUBMED    19740742
REFERENCE   2  (bases 1 to 5539)
  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: 
            "10.1074/jbc.M109.027995"; journalName: "J Biol Chem"; date: 
            "2009-10-30- 30"; volume: "284"; issue: "44"; pages: "30159-66"
COMMENT     Sequence Label: pCMV Flag WT-PTEN
FEATURES             Location/Qualifiers
     source          1..5539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        621..639
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      674..683
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             683..706
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1969..1985)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1970..1986)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2041..2060)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2042..2060)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2334..2455
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2371..2390)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2425..2444
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2462..2917)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2599..2620)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2811..2830
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2944..3048
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3050..3407
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3050..3241
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3073..3093)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3258..3393
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3442..4236
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3442..4236
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3496..3515)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4106..4125
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4424..4443)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4468..4515
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4844..5432
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5333..5352
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgattacg ccaagctcga aattaaccct cactaaaggg aacaaaagct ggagctccac
      661 cgcggtggcg gccgccacca tggattacaa ggatgacgac gataagagcc cgggcggatc
      721 catgacagcc atcatcaaag agatcgttag cagaaacaaa aggagatatc aagaggatgg
      781 attcgactta gacttgacct atatttatcc aaacattatt gctatgggat ttcctgcaga
      841 aagacttgaa ggcgtataca ggaacaatat tgatgatgta gtaaggtttt tggattcaaa
      901 gcataaaaac cattacaaga tatacaatct ttgtgctgaa agacattatg acaccgccaa
      961 atttaattgc agagttgcac aatatccttt tgaagaccat aacccaccac agctagaact
     1021 tatcaaaccc ttttgtgaag atcttgacca atggctaagt gaagatgaca atcatgttgc
     1081 agcaattcac tgtaaagctg gaaagggacg aactggtgta atgatatgtg catatttatt
     1141 acatcggggc aaatttttaa aggcacaaga ggccctagat ttctatgggg aagtaaggac
     1201 cagagacaaa aagggagtaa ctattcccag tcagaggcgc tatgtgtatt attatagcta
     1261 cctgttaaag aatcatctgg attatagacc agtggcactg ttgtttcaca agatgatgtt
     1321 tgaaactatt ccaatgttca gtggcggaac ttgcaatcct cagtttgtgg tctgccagct
     1381 aaaggtgaag atatattcct ccaattcagg acccacacga cgggaagaca agttcatgta
     1441 ctttgagttc cctcagccgt tacctgtgtg tggtgatatc aaagtagagt tcttccacaa
     1501 acagaacaag atgctaaaaa aggacaaaat gtttcacttt tgggtaaata cattcttcat
     1561 accaggacca gaggaaacct cagaaaaagt agaaaatgga agtctatgtg atcaagaaat
     1621 cgatagcatt tgcagtatag agcgtgcaga taatgacaag gaatatctag tacttacttt
     1681 aacaaaaaat gatcttgaca aagcaaataa agacaaagcc aaccgatact tttctccaaa
     1741 ttttaaggtg aagctgtact tcacaaaaac agtagaggag ccgtcaaatc cagaggctag
     1801 cagttcaact tctgtaacac cagatgttag tgacaatgaa cctgatcatt atagatattc
     1861 tgacaccact gactctgatc cagagaatga accttttgat gaagatcagc atacacaaat
     1921 tacaaaagtc tgaaagcttg ggaagggcga attcgatatc aagcttatcg ataccgtcga
     1981 cctcgagggg gggcccggta ccttaattaa ttaaggtacc aggtaagtgt acccaattcg
     2041 ccctatagtg agtcgtatta caattcactc gatcgccctt cccaacagtt gcgcagcctg
     2101 aatggcgaat ggagatccaa tttttaagtg tataatgtgt taaactactg attctaattg
     2161 tttgtgtatt ttagattcac agtcccaagg ctcatttcag gcccctcagt cctcacagtc
     2221 tgttcatgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     2281 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2341 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2401 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg
     2461 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     2521 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     2581 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     2641 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     2701 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2761 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2821 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2881 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     2941 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3001 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagaatcc tgaggcggaa
     3061 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3121 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3181 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3241 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3301 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3361 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     3421 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3481 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3541 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3601 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     3661 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3721 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3781 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3841 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3901 gatcaggatg atctggacga agaacatcag gggctcgcgc cagccgaact gttcgccagg
     3961 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4021 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4081 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agaacttggc
     4141 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4201 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4261 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4321 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4381 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     4441 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     4501 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     4561 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     4621 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     4681 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4741 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4801 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4861 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4921 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4981 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5041 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5101 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5161 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5221 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5281 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5341 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5401 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5461 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5521 tggataaccg tattaccgc
//
