LOCUS       sequence-349426-        6310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector encoding NDUFV3 and EmGFP-Bsd.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6310)
  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 6310)
  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: NDUFV3_pcDNA6.2/EmGFP-Bsd
FEATURES             Location/Qualifiers
     source          1..6310
                     /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(1032..1056)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1082..1123
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1163..1182)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1207..1255
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1457..1885
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1544..1563)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1754..1775
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1900..2399
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1935..1958)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2332..2351
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        2413..2460
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(2479..2481,2482..2484,2485..3195)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2524..2545)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2785..2804)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3132..3153
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3196..3591
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3749..3870
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3786..3805)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3840..3859
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3919..3935)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3919..3935)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3932..3954)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3943..3959
                     /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(3967..3973,3974..3991,3992..3997))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4012..4033
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4150..4167)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4321..4909)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4401..4420)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5080..5871,5872..5940))
                     /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     5703..5722
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5941..6045)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6120..6139)
                     /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 accatggctg ccccgtgttt
      721 gctgcggcaa ggacgagccg gggcgctgaa gactatgctc caggaagccc aggtgtttcg
      781 aggacttgct tctacggttt ctttgtctgc ggaatcaggg aagagtgaaa agggtcagcc
      841 acagaattcc aagaagcaaa gtccaccaaa aaagccagcc ccagtgcctg ctgagccgtt
      901 tgacaacact acctacaaga acctgcagca tcatgactac agcacgtaca ccttcttaga
      961 cctcaacctc gaactctcaa aattcaggat gcctcagccc tcctcaggcc gggagtcacc
     1021 tcgacactag aacccagctt tcttgtacaa agtggttgat ctagagggcc cgcggttcga
     1081 aggtaagcct atccctaacc ctctcctcgg tctcgattct acgcgtaccg gttagtaatg
     1141 agtttaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     1201 ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa
     1261 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     1321 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     1381 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc
     1441 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     1501 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     1561 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     1621 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     1681 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     1741 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     1801 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     1861 atttaacaaa aatttaacgc gaattatcct cgagctaccg ggtaggggag gcgcttttcc
     1921 caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg
     1981 gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg
     2041 gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc
     2101 agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga
     2161 tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc
     2221 tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca
     2281 ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca
     2341 cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga
     2401 ctctagacac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac
     2461 aaggtgagga actaaaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     2521 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     2581 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     2641 cccgtgccct ggcccaccct cgtgaccacc ttcacctacg gcgtgcagtg cttcgcccgc
     2701 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2761 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     2821 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2881 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaaggt ctatatcacc
     2941 gccgacaagc agaagaacgg catcaaggtg aacttcaaga cccgccacaa catcgaggac
     3001 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     3061 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     3121 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     3181 gacgagctgt acaaggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca
     3241 acggctacaa tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc
     3301 tctagcgacg gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt
     3361 gcagaactcg tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc
     3421 gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg
     3481 cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg
     3541 gcagttggga ttcgtgaatt gctgccctct ggttatgtgt gggagggcta agcacttcgt
     3601 ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga
     3661 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     3721 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     3781 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3841 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     3901 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3961 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4021 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4081 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4141 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4201 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4261 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4321 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4381 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4441 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4501 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4561 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4621 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4681 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4741 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4801 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4861 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4921 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4981 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5041 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5101 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5161 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5221 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5281 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5341 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5401 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agcttcggtt cccaacgatc
     5461 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5521 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5581 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5641 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5701 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5761 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5821 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5881 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5941 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6001 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6061 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     6121 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     6181 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     6241 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     6301 tatacgcgtt
//
