LOCUS       sequence-343000-        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"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    911..935
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1266..1290)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1316..1357
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1397..1416)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1441..1489
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1691..2119
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1778..1797)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1988..2009
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2134..2633
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2169..2192)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2566..2585
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        2647..2694
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(2713..2715,2716..2718,2719..3429)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2758..2779)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3019..3038)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3366..3387
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3430..3825
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3983..4104
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4020..4039)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4074..4093
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4153..4169)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4153..4169)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4166..4188)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4177..4193
                     /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(4201..4207,4208..4225,4226..4231))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4246..4267
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4384..4401)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4555..5143)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4635..4654)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5314..6105,6106..6174))
                     /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     5937..5956
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6175..6279)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagctatca acaagtttgt acaaaaaagc aggcaccatg gctgccccgt gtttgctgcg
      961 gcaaggacga gccggggcgc tgaagactat gctccaggaa gcccaggtgt ttcgaggact
     1021 tgcttctacg gtttctttgt ctgcggaatc agggaagagt gaaaagggtc agccacagaa
     1081 ttccaagaag caaagtccac caaaaaagcc agccccagtg cctgctgagc cgtttgacaa
     1141 cactacctac aagaacctgc agcatcatga ctacagcacg tacaccttct tagacctcaa
     1201 cctcgaactc tcaaaattca ggatgcctca gccctcctca ggccgggagt cacctcgaca
     1261 ctagtaccca gctttcttgt acaaagtggt tgatctagag ggcccgcggt tcgaaggtaa
     1321 gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggttagt aatgagttta
     1381 aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     1441 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg
     1501 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     1561 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     1621 gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg
     1681 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     1741 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     1801 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     1861 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     1921 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     1981 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2041 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2101 caaaaattta acgcgaatta tcctcgagct accgggtagg ggaggcgctt ttcccaaggc
     2161 agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct
     2221 ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc
     2281 ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg
     2341 cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca
     2401 gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct
     2461 ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg
     2521 ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc
     2581 aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgactctag
     2641 acacgtgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
     2701 aggaactaaa ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2761 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     2821 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2881 ccctggccca ccctcgtgac caccttcacc tacggcgtgc agtgcttcgc ccgctacccc
     2941 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3001 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3061 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3121 atcctggggc acaagctgga gtacaactac aacagccaca aggtctatat caccgccgac
     3181 aagcagaaga acggcatcaa ggtgaacttc aagacccgcc acaacatcga ggacggcagc
     3241 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     3301 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     3361 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3421 ctgtacaagg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct
     3481 acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc
     3541 gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa
     3601 ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg
     3661 atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc
     3721 gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt
     3781 gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctaagcact tcgtggccga
     3841 ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt
     3901 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     3961 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     4021 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4081 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     4141 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4201 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4261 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4321 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4381 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4441 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4501 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4561 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4621 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4681 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4741 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4801 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4861 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4921 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4981 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5041 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5101 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5161 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5221 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5281 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5341 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5401 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5461 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5521 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5581 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5641 ggtgtcacgc tcgtcgtttg gtatggcttc attcagcttc ggttcccaac gatcaaggcg
     5701 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5761 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5821 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5881 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5941 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6001 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6061 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6121 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6181 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6241 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6301 acctgacgtc
//
