LOCUS       sequence-349424-        7285 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector encoding PRMT2 and EmGFP-Bsd.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7285)
  AUTHORS   Moyer AJ, Fernandez FX, Li Y, Klinedinst DK, Florea LD, Kazuki Y, 
            Oshimura M, Reeves RH
  TITLE     Overexpression screen of chromosome 21 genes reveals modulators of 
            Sonic hedgehog signaling relevant to Down syndrome.
  JOURNAL   Dis Model Mech. 2023 Apr 1;16(4):dmm049712. doi: 10.1242/dmm.049712.
            Epub 2023 Apr 13.
  PUBMED    36995257
REFERENCE   2  (bases 1 to 7285)
  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: "Dis Model 
            Mech."; date: "2023-04-1"; volume: "16(4):dmm049712. doi"; pages: " 
            10.1242/dmm.049712. Epub 2023 Apr 13"
COMMENT     Sequence Label: PRMT2_pcDNA6.2/EmGFP-Bsd
FEATURES             Location/Qualifiers
     source          1..7285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     249..268
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        249..267
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    297..321
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1627..1651)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1677..1718
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1758..1777)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1802..1850
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2052..2480
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2139..2158)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2349..2370
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2495..2994
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2530..2553)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2927..2946
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        3008..3055
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(3074..3076,3077..3079,3080..3790)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3119..3140)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3380..3399)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3727..3748
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3791..4186
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4344..4465
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4381..4400)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4435..4454
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4514..4530)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4514..4530)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4527..4549)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4538..4554
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4562..4568,4569..4586,4587..4592))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4607..4628
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4745..4762)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4916..5504)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4996..5015)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5675..6466,6467..6535))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPKLNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6298..6317
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6536..6640)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6715..6734)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6906..7285
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      241 atcgaaatta atacgactca ctatagggag acccaagctg gctagttaag ctatcaacaa
      301 gtttgtacaa aaaagcaggc accatggcaa catcaggtga ctgtcccaga agtgaatcgc
      361 agggagaaga gcctgctgag tgcagtgagg cgggtctcct gcaggaggga gtacagccag
      421 aggagtttgt ggccatcgcg gactacgctg ccaccgatga gacccagctc agttttttga
      481 gaggagaaaa aattcttatc ctgagacaaa ccactgcaga ttggtggtgg ggtgagcgtg
      541 cgggctgctg tgggtacatt ccggcaaacc atgtggggaa gcacgtggat gagtacgacc
      601 ccgaggacac gtggcaggat gaagagtact tcggcagcta tggaactctg aaactccact
      661 tggagatgtt ggcagaccag ccacgaacaa ctaaatacca cagtgtcatc ctgcagaata
      721 aagaatccct gacggataaa gtcatcctgg acgtgggctg tgggactggg atcatcagtc
      781 tcttctgtgc acactatgcg cggcctagag cggtgtacgc ggtggaggcc agtgagatgg
      841 cacagcacac ggggcagctg gtcttgcaga acggctttgc tgacatcatc accgtgtacc
      901 agcagaaggt ggaggatgtg gtgctgcccg agaaggtgga cgtgctggtg tctgagtgga
      961 tggggacctg cctgctgttt gagttcatga tcgagtccat cctgtatgcc cgggatgcct
     1021 ggctgaagga ggacggggtc atttggccca ccatggctgc gttgcacctt gtgccctgca
     1081 gtgctgataa ggattatcgt agcaaggtgc tcttctggga caacgcgtac gagttcaacc
     1141 tcagcgctct gaaatcttta gcagttaagg agtttttttc aaagcccaag tataaccaca
     1201 ttttgaaacc agaagactgt ctctctgaac cgtgcactat attgcagttg gacatgagaa
     1261 ccgtgcaaat ttctgatcta gagaccctga ggggcgagct gcgcttcgac atcaggaagg
     1321 cggggaccct gcacggcttc acggcctggt ttagcgtcca cttccagagc ctgcaggagg
     1381 ggcagccgcc gcaggtgctc agcaccgggc ccttccaccc caccacacac tggaagcaga
     1441 cgctgttcat gatggacgac ccagtccctg tccatacagg agacgtggtc acgggttcag
     1501 ttgtgttgca gagaaaccca gtgtggataa ggcacatgtc tgtggctctg agctgggctg
     1561 tcacttccag acaagacccc acatctcaaa aagttggaga aaaagtcttc cccatctgga
     1621 gatagaaccc agctttcttg tacaaagtgg ttgatctaga gggcccgcgg ttcgaaggta
     1681 agcctatccc taaccctctc ctcggtctcg attctacgcg taccggttag taatgagttt
     1741 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     1801 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     1861 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     1921 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     1981 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     2041 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2101 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     2161 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     2221 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     2281 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2341 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     2401 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     2461 acaaaaattt aacgcgaatt atcctcgagc taccgggtag gggaggcgct tttcccaagg
     2521 cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc
     2581 tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc
     2641 cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc
     2701 gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac
     2761 agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc
     2821 tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc
     2881 gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt
     2941 caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct ttcgactcta
     3001 gacacgtgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     3061 gaggaactaa accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     3121 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     3181 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     3241 gccctggccc accctcgtga ccaccttcac ctacggcgtg cagtgcttcg cccgctaccc
     3301 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     3361 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     3421 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     3481 catcctgggg cacaagctgg agtacaacta caacagccac aaggtctata tcaccgccga
     3541 caagcagaag aacggcatca aggtgaactt caagacccgc cacaacatcg aggacggcag
     3601 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     3661 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     3721 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     3781 gctgtacaag gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc
     3841 tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag
     3901 cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga
     3961 actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc
     4021 gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct
     4081 cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt
     4141 tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaagcac ttcgtggccg
     4201 aggagcagga ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt
     4261 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4321 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     4381 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4441 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     4501 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4561 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4621 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4681 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4741 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4801 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4861 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4921 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4981 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5041 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5101 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5161 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5221 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5281 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5341 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5401 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5461 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5521 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5581 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5641 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5701 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5761 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5821 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5881 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5941 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6001 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctt cggttcccaa cgatcaaggc
     6061 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6121 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6181 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6241 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6301 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6361 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6421 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6481 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6541 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6601 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     6661 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     6721 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     6781 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     6841 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
     6901 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     6961 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     7021 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     7081 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     7141 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     7201 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     7261 cgtattagtc atcgctatta ccatg
//
