LOCUS       sequence-349420-        7110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector encoding CFAP410 and EmGFP-Bsd.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7110)
  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 7110)
  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: CFAP410_pcDNA6.2/EmGFP-Bsd
FEATURES             Location/Qualifiers
     source          1..7110
                     /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(1833..1857)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1883..1924
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1964..1983)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2008..2056
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2258..2686
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2345..2364)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2555..2576
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2701..3199
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2736..2759)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3132..3151
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        3213..3260
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(3279..3281,3282..3284,3285..3995)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3324..3345)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3585..3604)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3932..3953
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3996..4391
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4549..4670
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4586..4605)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4640..4659
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4719..4735)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4719..4735)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4732..4754)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4743..4759
                     /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(4767..4773,4774..4791,4792..4797))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4812..4833
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4950..4967)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5121..5709)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5201..5220)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5880..6671,6672..6740))
                     /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     6503..6522
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6741..6845)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6920..6939)
                     /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 accatgaagc tgacgcggaa
      721 gatggttctg acccgggcca aggcctcgga gctgcacagc gtgcgcaagc tcaactgctg
      781 gggcagccgc ctcacagata tctccatttg ccaggagatg cccagcctgg aggtgatcac
      841 gctcagtgtc aacagcatct ccaccctgga gcctgtgagc cggtgccagc gcctgagtga
      901 gctgtacctg cggaggaacc gcatccccag cctggctgag ctcttctacc tgaaggggct
      961 gccgcgtctg cgggtgctgt ggctggccga gaacccgtgc tgcggcacca gcccccaccg
     1021 ctaccgcatg accgtgctgc gcaccctgcc gcgcctacag aagctggaca accaggctgt
     1081 gacggaggag gagctgtccc gtgcactgag tgagggagag gagatcactg cggccccaga
     1141 gagagagggc acaggccacg gcggccccaa gctatgctgc acactgagct ccctcagctc
     1201 cgctgctgag actggccggg acccgctgga cagcgaggag gaggcaaccg gcgcccagga
     1261 tgaacgtggc ctgaagccgc cttcccgggg ccagtttcct tccctctcag ccagggatgc
     1321 ctcgagcagc cacaggggca gggtgagtgg cgggccgcta ggggccgcgg ctgcctctgc
     1381 ccactgcacc cactgcacag aaaccgtggg gagggagcat ggagcctcac agggccccgt
     1441 ggggagggag catggagcct cacagggcct tgaagagctg tgccccaggg ggagctgcgt
     1501 gtgcgggtct gtgaatgcgc acacacgtgt aacacgtgcc ccgcacggag ccgtcctggc
     1561 ccctcagcct ctcctgctgt cctggtctgt ggaatgtggg cccgggccct gctgggctga
     1621 gggcaacagg agtcacgtgg aagaggtgcc acacacgcgt ccacaggcgg ggctcctctg
     1681 ctcagattct ccgagtgtgc cgaacgtcct gactgccatc ctgctgctgc tgcgggagct
     1741 ggatgcagag gggctggagg ccgtgcagca gactgtgggc agccggctgc aggccctgcg
     1801 tggggaagag gtgcaggagc acgccgagta gaacccagct ttcttgtaca aagtggttga
     1861 tctagagggc ccgcggttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc
     1921 tacgcgtacc ggttagtaat gagtttaaac gggggaggct aactgaaaca cggaaggaga
     1981 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt
     2041 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     2101 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     2161 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     2221 gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     2281 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2341 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2401 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2461 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2521 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2581 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2641 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattatcc tcgagctacc
     2701 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg
     2761 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc
     2821 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca
     2881 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac
     2941 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt
     3001 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg
     3061 gtgggtccgg gggcgggctc aggggcggct caggggcggg gcgggcgccc gaaggtcctc
     3121 cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc
     3181 ctcatctccg ggcctttcga ctctagacac gtgttgacaa ttaatcatcg gcatagtata
     3241 tcggcatagt ataatacgac aaggtgagga actaaaccat ggtgagcaag ggcgaggagc
     3301 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     3361 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     3421 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcacctacg
     3481 gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     3541 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     3601 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     3661 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     3721 gccacaaggt ctatatcacc gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     3781 cccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     3841 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     3901 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     3961 ccgggatcac tctcggcatg gacgagctgt acaaggccaa gcctttgtct caagaagaat
     4021 ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca
     4081 gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc
     4141 attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag
     4201 ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct
     4261 gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg
     4321 acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt
     4381 gggagggcta agcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat
     4441 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4501 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4561 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4621 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4681 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4741 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4801 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4861 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4921 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4981 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5041 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5101 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5161 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5221 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5281 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5341 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5401 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5461 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5521 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5581 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5641 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5701 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5761 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5821 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5881 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5941 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6001 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6061 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6121 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6181 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6241 agcttcggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6301 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6361 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6421 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6481 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6541 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6601 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6661 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6721 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6781 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6841 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     6901 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     6961 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     7021 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     7081 ttcgcgatgt acgggccaga tatacgcgtt
//
