LOCUS       sequence-349704-        6295 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector encoding SUMO3 and EmGFP-Bsd.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6295)
  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 6295)
  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: SUMO3_pcDNA6.2/EmGFP-Bsd
FEATURES             Location/Qualifiers
     source          1..6295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    677..701
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             704..979
                     /codon_start=1
                     /product="human small ubiquitin-related modifier 3"
                     /label=SUMO3
                     /translation="MSEEKPKEGVKTENDHINLKVAGQDGSVVQFKIKRHTPLSKLMKA
                     YCERQGLSMRQIRFRFDGQPINETDTPAQLEMEDEDTIDVFQQQTGG"
     protein_bind    complement(1017..1041)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1067..1108
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1148..1167)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1192..1240
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1442..1870
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1529..1548)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1739..1760
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1885..2384
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1920..1943)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2317..2336
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        2398..2445
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(2464..2466,2467..2469,2470..3180)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2509..2530)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2770..2789)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3117..3138
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3181..3576
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3734..3855
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3771..3790)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3825..3844
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3904..3920)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3904..3920)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3917..3939)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3928..3944
                     /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(3952..3958,3959..3976,3977..3982))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3997..4018
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4135..4152)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4306..4894)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4386..4405)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5065..5856,5857..5925))
                     /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     5688..5707
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5926..6030)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6105..6124)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaag ctatcaacaa gtttgtacaa aaaagcaggc accatgtccg aggagaagcc
      721 caaggagggt gtgaagacag agaatgacca catcaacctg aaggtggccg ggcaggacgg
      781 ctccgtggtg cagttcaaga tcaagaggca cacgccgctg agcaagctga tgaaggccta
      841 ctgcgagagg cagggcttgt caatgaggca gatcagattc aggttcgacg ggcagccaat
      901 caatgaaact gacactccag cacagctgga gatggaggac gaggacacca tcgacgtgtt
      961 ccagcagcag acgggaggtg tgccggagag cagcctggca gggcacagtt tctagaaccc
     1021 agctttcttg tacaaagtgg ttgatctaga gggcccgcgg ttcgaaggta agcctatccc
     1081 taaccctctc ctcggtctcg attctacgcg taccggttag taatgagttt aaacggggga
     1141 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     1201 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     1261 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     1321 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     1381 gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc
     1441 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     1501 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     1561 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     1621 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     1681 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     1741 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     1801 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     1861 aacgcgaatt atcctcgagc taccgggtag gggaggcgct tttcccaagg cagtctggag
     1921 catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca
     1981 cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc
     2041 caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca
     2101 ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg
     2161 agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt
     2221 tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg
     2281 gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca
     2341 cgtctgccgc gctgttctcc tcttcctcat ctccgggcct ttcgactcta gacacgtgtt
     2401 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
     2461 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     2521 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     2581 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     2641 accctcgtga ccaccttcac ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg
     2701 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2761 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2821 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2881 cacaagctgg agtacaacta caacagccac aaggtctata tcaccgccga caagcagaag
     2941 aacggcatca aggtgaactt caagacccgc cacaacatcg aggacggcag cgtgcagctc
     3001 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     3061 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     3121 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     3181 gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac
     3241 agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc
     3301 atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg
     3361 ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat
     3421 gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat
     3481 cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt
     3541 gaattgctgc cctctggtta tgtgtgggag ggctaagcac ttcgtggccg aggagcagga
     3601 ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     3661 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     3721 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3781 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3841 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     3901 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3961 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4021 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4081 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4141 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4201 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4261 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4321 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4381 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4441 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4501 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4561 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4621 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4681 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4741 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4801 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4861 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4921 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4981 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5041 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5101 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5161 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5221 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5281 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5341 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5401 ctcgtcgttt ggtatggctt cattcagctt cggttcccaa cgatcaaggc gagttacatg
     5461 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5521 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5581 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5641 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5701 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5761 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5821 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5881 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5941 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6001 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6061 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
     6121 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     6181 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     6241 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgtt
//
