LOCUS       sequence-142546-        7134 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 7134)
  AUTHORS   Kolli S, Zito CI, Mossink MH, Wiemer EA, Bennett AM
  TITLE     The major vault protein is a novel substrate for the tyrosine 
            phosphatase SHP-2 and scaffold protein in epidermal growth factor 
            signaling.
  JOURNAL   J Biol Chem. 2004 Jul 9. 279(28):29374-85.
  PUBMED    15133037
REFERENCE   2  (bases 1 to 7134)
  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"; journalName: "J Biol 
            Chem. 2004 Jul 9. 279(28):29374-85."
COMMENT     Sequence Label: pIRES2-EGFP-SHP2DA
FEATURES             Location/Qualifiers
     source          1..7134
                     /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"
     misc_feature    join(2500..3074,3075..3077,3078..3086)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2667..2684)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2894..2913
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3087..3089,3090..3092,3093..3806)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3087..3089,3090..3092,3093..3803)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3132..3153)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3393..3412)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3740..3761
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3929..4050
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3966..3985)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4020..4039
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4057..4512)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4194..4215)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4406..4425
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4539..4643
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4645..5002
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4645..4836
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4668..4688)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4853..4988
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5037..5831
                     /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             5037..5831
                     /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(5091..5110)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5701..5720
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6019..6038)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6063..6110
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6439..7027
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6928..6947
                     /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 agcgctaccg gactcagatc tcgagctcaa gcttcgaatt ccgggcggga ggaacatgac
      661 atcgcggaga tggtttcacc caaatatcac tggtgtggag gcagaaaacc tactgttgac
      721 aagaggagtt gatggcagtt ttttggcaag gcctagtaaa agtaaccctg gagacttcac
      781 actttccgtt agaagaaatg gagctgtcac ccacatcaag attcagaaca ctggtgatta
      841 ctatgacctg tatggagggg agaaatttgc cactttggct gagttggtcc agtattacat
      901 ggaacatcac gggcaattaa aagagaagaa tggagatgtc attgagctta aatatcctct
      961 gaactgtgca gatcctacct ctgaaaggtg gtttcatgga catctctctg ggaaagaagc
     1021 agagaaatta ttaactgaaa aaggaaaaca tggtagtttt cttgtacgag agagccagag
     1081 ccaccctgga gattttgttc tttctgtgcg cactggtgat gacaaagggg agagcaatga
     1141 cggcaagtct aaagtgaccc atgttatgat tcgctgtcag gaactgaaat acgacgttgg
     1201 tggaggagaa cggtttgatt ctttgacaga tcttgtggaa cattataaga agaatcctat
     1261 ggtggaaaca ttgggtacag tactacaact caagcagccc cttaacacga ctcgtataaa
     1321 tgctgctgaa atagaaagca gagttcgaga actaagcaaa ttagctgaga ccacagataa
     1381 agtcaaacaa ggcttttggg aagaatttga gacactacaa caacaggagt gcaaacttct
     1441 ctacagccga aaagagggtc aaaggcaaga aaacaaaaac aaaaatagat ataaaaacat
     1501 cctgcccttt gatcatacca gggttgtcct acacgatggt gatcccaatg agcctgtttc
     1561 agattacatc aatgcaaata tcatcatgcc tgaatttgaa accaagtgca acaattcaaa
     1621 gcccaaaaag agttacattg ccacacaagg ctgcctgcaa aacacggtga atgacttttg
     1681 gcggatggtg ttccaagaaa actcccgagt gattgtcatg acaacgaaag aagtggagag
     1741 aggaaagagt aaatgtgtca aatactggcc tgatgagtat gctctaaaag aatatggcgt
     1801 catgcgtgtt aggaacgtca aagaaagcgc cgctcatgac tatacgctaa gagaacttaa
     1861 actttcaaag gttggacaag ggaatacgga gagaacggtc tggcaatacc actttcggac
     1921 ctggccggcc cacggcgtgc ccagcgaccc tgggggcgtg ctggacttcc tggaggaggt
     1981 gcaccataag caggagagca tcatggatgc agggccggtc gtggtgcact gcagtgctgg
     2041 aattggccgg acagggacgt tcattgtgat tgatattctt attgacatca tcagagagaa
     2101 aggtgttgac tgcgatattg acgttcccaa aaccatccag atggtgcggt ctcagaggtc
     2161 agggatggtc gcgacagaag cacagtaccg atttatctat atggcggtcc agcattatat
     2221 tgaaacacta cagcgcagga ttgaagaaga gcagaaaagc aagaggaaag ggcacgaata
     2281 tacaaatatt aagtattctc tagcggacca gacgagtgga gatcagagcc ctctcccgcc
     2341 ttgtactcca acgccaccct gtgcagaaat gagagaagac agtgctagag tctatgaaaa
     2401 cgtgggcctg atgcaacagc agaaaagttt cagaaaactc atctctgaag aggatctgtg
     2461 agaattctgc agtcgacggt accgcgggcc cgggatccgc ccctctccct cccccccccc
     2521 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     2581 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     2641 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     2701 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     2761 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     2821 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     2881 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     2941 ggtaccccat tgtatgggat ctgatctggg gcctcggtac acatgcttta catgtgttta
     3001 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     3061 aacacgatga taatatggcc acaaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     3121 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     3181 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     3241 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     3301 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     3361 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     3421 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     3481 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     3541 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     3601 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     3661 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     3721 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     3781 tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac
     3841 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     3901 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     3961 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4021 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     4081 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4141 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4201 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4261 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4321 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4381 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4441 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4501 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4561 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4621 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     4681 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4741 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4801 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4861 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4921 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4981 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     5041 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5101 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5161 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     5221 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5281 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5341 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5401 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5461 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5521 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     5581 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5641 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5701 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5761 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5821 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5881 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5941 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6001 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     6061 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     6121 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     6181 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     6241 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     6301 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6361 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6421 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6481 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6541 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6601 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6661 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6721 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6781 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6841 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6901 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6961 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7021 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7081 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgc
//
