LOCUS       sequence-349410-        6988 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector encoding HSF2BP and EmGFP-Bsd.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6988)
  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 6988)
  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: HSF2BP_pcDNA6.2/EmGFP-Bsd
FEATURES             Location/Qualifiers
     source          1..6988
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     249..268
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        249..267
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    297..321
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1330..1354)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1380..1421
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1461..1480)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1505..1553
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1755..2183
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1842..1861)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2052..2073
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2198..2697
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2233..2256)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2630..2649
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        2711..2758
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(2777..2779,2780..2782,2783..3493)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2822..2843)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3083..3102)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3430..3451
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3494..3889
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4047..4168
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4084..4103)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4138..4157
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4217..4233)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4217..4233)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4230..4252)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4241..4257
                     /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(4265..4271,4272..4289,4290..4295))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4310..4331
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4448..4465)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4619..5207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4699..4718)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5378..6169,6170..6238))
                     /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     6001..6020
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6239..6343)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6418..6437)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6609..6988
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      241 atcgaaatta atacgactca ctatagggag acccaagctg gctagttaag ctatcaacaa
      301 gtttgtacaa aaaagcaggc accatgggcg aagcgggcgc cgctgaggag gcctgccggc
      361 acatgggaac taaagaggaa tttgttaaag tcagaaagaa ggatctggaa cggctgacaa
      421 ctgaagtgat gcaaatacgg gacttcttac ccagaatact aaatggggag gtgctggaga
      481 gcttccagaa attaaagatt gtagaaaaaa acctggaaag gaaagagcaa gaattagagc
      541 agctgaaaat ggattgtgag cactttaaag cccgcctgga aaccgtgcag gccgacaaca
      601 taagagagaa gaaggagaaa ctggctcttc gacagcagtt gaatgaagcg aagcagcaac
      661 tcctgcagca ggcagagtat tgtacagaaa tgggagcagc agcgtgtacc ctcttgtggg
      721 gtgtctccag cagtgaggaa gtcgtcaagg ccattttggg aggagataaa gctttgaagt
      781 ttttcagcat cactggtcaa acaatggaga gttttgtgaa gtcgttagac ggtgatgtcc
      841 aggagctgga ttcggatgaa agtcagtttg ttttcgctct ggctggaatt gtcacgaatg
      901 ttgctgctat agcatgtggt cgtgaattct tggttaattc aagccgggtg ctcttggaca
      961 ccatattgca gcttctggga gacttgaagc caggacagtg taccaaactc aaagtgctaa
     1021 tgctgatgtc cctatacaat gtaagcatca atttgaaagg cttgaaatac atcagcgaga
     1081 gtccaggatt catccctttg ctgtggtggc ttttgagtga tccagatgca gaggtgtgcc
     1141 ttcatgtact gaggcttgtc cagtctgtgg ttctggaacc tgaagtcttc tccaagtcgg
     1201 cctctgagtt ccggagctcc ctgcccctgc aacgcatcct ggcaatgtcc aagagccgca
     1261 acccccgcct gcaaaccgca gcccaggagc tcctggaaga tctccgcact ctggagcata
     1321 atgtgtagaa cccagctttc ttgtacaaag tggttgatct agagggcccg cggttcgaag
     1381 gtaagcctat ccctaaccct ctcctcggtc tcgattctac gcgtaccggt tagtaatgag
     1441 tttaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     1501 atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg
     1561 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     1621 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     1681 gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc
     1741 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     1801 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     1861 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     1921 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     1981 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2041 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     2101 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     2161 ttaacaaaaa tttaacgcga attatcctcg agctaccggg taggggaggc gcttttccca
     2221 aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc
     2281 ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt
     2341 ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag
     2401 ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg
     2461 gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt
     2521 tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg
     2581 ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg
     2641 cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgact
     2701 ctagacacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     2761 ggtgaggaac taaaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     2821 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     2881 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     2941 cgtgccctgg cccaccctcg tgaccacctt cacctacggc gtgcagtgct tcgcccgcta
     3001 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     3061 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     3121 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     3181 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaaggtct atatcaccgc
     3241 cgacaagcag aagaacggca tcaaggtgaa cttcaagacc cgccacaaca tcgaggacgg
     3301 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     3361 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     3421 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     3481 cgagctgtac aaggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac
     3541 ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc
     3601 tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc
     3661 agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt
     3721 cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct
     3781 tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc
     3841 agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag cacttcgtgg
     3901 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
     3961 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4021 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4081 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4141 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     4201 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4261 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4321 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4381 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4441 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4501 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4561 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4621 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4681 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4741 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4801 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4861 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4921 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4981 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5041 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5101 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5161 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5221 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5281 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5341 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5401 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5461 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5521 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5581 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5641 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5701 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag cttcggttcc caacgatcaa
     5761 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5821 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5881 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5941 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6001 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6061 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6121 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6181 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6241 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6301 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6361 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     6421 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     6481 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     6541 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     6601 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     6661 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     6721 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     6781 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     6841 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     6901 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     6961 ctacgtatta gtcatcgcta ttaccatg
//
