LOCUS       sequence-349698-        6286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector encoding HMGN1 and EmGFP-Bsd.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6286)
  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 6286)
  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: HMGN1_pcDNA6.2/EmGFP-Bsd
FEATURES             Location/Qualifiers
     source          1..6286
                     /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(1008..1032)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1058..1099
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1139..1158)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1183..1231
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1433..1861
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1520..1539)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1730..1751
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1876..2375
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1911..1934)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2308..2327
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        2389..2436
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(2455..2457,2458..2460,2461..3171)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2500..2521)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2761..2780)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3108..3129
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3172..3567
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3725..3846
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3762..3781)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3816..3835
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3895..3911)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3895..3911)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3908..3930)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3919..3935
                     /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(3943..3949,3950..3967,3968..3973))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3988..4009
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4126..4143)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4297..4885)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4377..4396)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5056..5847,5848..5916))
                     /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     5679..5698
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5917..6021)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6096..6115)
                     /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 accatgccca agaggaaggt
      721 cagctccgcc gaaggcgccg ccaaggaaga gcccaagagg agatcggcgc ggttgtcagc
      781 taaacctcct gcaaaagtgg aagcgaagcc gaaaaaggca gcagcgaagg ataaatcttc
      841 agacaaaaaa gtgcaaacaa aagggaaaag gggagcaaag ggaaaacagg ccgaagtggc
      901 taaccaagaa actaaagaag acttacctgc ggaaaacggg gaaacgaaga ctgaggagag
      961 tccagcctct gatgaagcag gagagaaaga agccaagtct gattagaacc cagctttctt
     1021 gtacaaagtg gttgatctag agggcccgcg gttcgaaggt aagcctatcc ctaaccctct
     1081 cctcggtctc gattctacgc gtaccggtta gtaatgagtt taaacggggg aggctaactg
     1141 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     1201 ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     1261 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     1321 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     1381 gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc
     1441 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1501 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     1561 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     1621 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     1681 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     1741 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     1801 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     1861 tatcctcgag ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt
     1921 tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca
     1981 catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta
     2041 ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac
     2101 aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga
     2161 agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc
     2221 agaggctggg aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg
     2281 gcgcccgaag gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg
     2341 cgctgttctc ctcttcctca tctccgggcc tttcgactct agacacgtgt tgacaattaa
     2401 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggtg
     2461 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2521 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     2581 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2641 accaccttca cctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
     2701 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2761 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2821 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2881 gagtacaact acaacagcca caaggtctat atcaccgccg acaagcagaa gaacggcatc
     2941 aaggtgaact tcaagacccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     3001 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     3061 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     3121 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa ggccaagcct
     3181 ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc
     3241 atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact
     3301 ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact
     3361 gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg
     3421 ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc
     3481 aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg
     3541 ccctctggtt atgtgtggga gggctaagca cttcgtggcc gaggagcagg actgacacgt
     3601 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3661 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3721 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3781 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3841 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     3901 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3961 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4021 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4081 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4141 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4201 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4261 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4321 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4381 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4441 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4501 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4561 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4621 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4681 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4741 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4801 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4861 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4921 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4981 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5041 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5101 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5161 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5221 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5281 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5341 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5401 tggtatggct tcattcagct tcggttccca acgatcaagg cgagttacat gatcccccat
     5461 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5521 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5581 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5641 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5701 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5761 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5821 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5881 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5941 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6001 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     6061 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     6121 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     6181 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
     6241 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgtt
//
