LOCUS       sequence-349723-        6448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector encoding SOD1 and EmGFP-Bsd.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6448)
  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 6448)
  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: SOD1_pcDNA6.2/EmGFP-Bsd
FEATURES             Location/Qualifiers
     source          1..6448
                     /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"
     protein_bind    complement(1170..1194)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1220..1261
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1301..1320)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1345..1393
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1595..2023
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1682..1701)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1892..1913
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2038..2537
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2073..2096)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2470..2489
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        2551..2598
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(2617..2619,2620..2622,2623..3333)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2662..2683)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2923..2942)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3270..3291
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3334..3729
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3887..4008
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3924..3943)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3978..3997
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4057..4073)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4057..4073)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4070..4092)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4081..4097
                     /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(4105..4111,4112..4129,4130..4135))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4150..4171
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4288..4305)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4459..5047)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4539..4558)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5218..6009,6010..6078))
                     /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     5841..5860
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6079..6183)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6258..6277)
                     /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 accatggcga cgaaggccgt
      721 gtgcgtgctg aagggcgacg gcccagtgca gggcatcatc aatttcgagc agaaggaaag
      781 taatggacca gtgaaggtgt ggggaagcat taaaggactg actgaaggcc tgcatggatt
      841 ccatgttcat gagtttggag ataatacagc aggctgtacc agtgcaggtc ctcactttaa
      901 tcctctatcc agaaaacacg gtgggccaaa ggatgaagag aggcatgttg gagacttggg
      961 caatgtgact gctgacaaag atggtgtggc cgatgtgtct attgaagatt ctgtgatctc
     1021 actctcagga gaccattgca tcattggccg cacactggtg gtccatgaaa aagcagatga
     1081 cttgggcaaa ggtggaaatg aagaaagtac aaagacagga aacgctggaa gtcgtttggc
     1141 ttgtggtgta attgggatcg cccaatagaa cccagctttc ttgtacaaag tggttgatct
     1201 agagggcccg cggttcgaag gtaagcctat ccctaaccct ctcctcggtc tcgattctac
     1261 gcgtaccggt tagtaatgag tttaaacggg ggaggctaac tgaaacacgg aaggagacaa
     1321 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt
     1381 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     1441 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     1501 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca
     1561 gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     1621 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     1681 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     1741 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     1801 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     1861 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     1921 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     1981 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attatcctcg agctaccggg
     2041 taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca
     2101 cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc
     2161 aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga
     2221 agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc
     2281 tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg
     2341 gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg
     2401 ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg
     2461 gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct
     2521 catctccggg cctttcgact ctagacacgt gttgacaatt aatcatcggc atagtatatc
     2581 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg tgagcaaggg cgaggagctg
     2641 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     2701 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     2761 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cacctacggc
     2821 gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     2881 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     2941 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     3001 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     3061 cacaaggtct atatcaccgc cgacaagcag aagaacggca tcaaggtgaa cttcaagacc
     3121 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     3181 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     3241 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     3301 gggatcactc tcggcatgga cgagctgtac aaggccaagc ctttgtctca agaagaatcc
     3361 accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc
     3421 gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat
     3481 tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct
     3541 ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc
     3601 ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac
     3661 agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg
     3721 gagggctaag cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc
     3781 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     3841 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     3901 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3961 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4021 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4081 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4141 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4201 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4261 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4321 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4381 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4441 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4501 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4561 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4621 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4681 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4741 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4801 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4861 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4921 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4981 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5041 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5101 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5161 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5221 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5281 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5341 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5401 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5461 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5521 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5581 cttcggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5641 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5701 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5761 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5821 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5881 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5941 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6001 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6061 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6121 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6181 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     6241 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     6301 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     6361 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     6421 cgcgatgtac gggccagata tacgcgtt
//
