LOCUS       sequence-349694-        8008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector encoding ABCG1 and EmGFP-Bsd.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8008)
  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 8008)
  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: ABCG1_pcDNA6.2/EmGFP-Bsd
FEATURES             Location/Qualifiers
     source          1..8008
                     /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(2362..2386)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2412..2453
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2493..2512)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2537..2585
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2787..3215
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2874..2893)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3084..3105
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3230..3717
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3265..3288)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3650..3669
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        3731..3778
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(3797..3799,3800..3802,3803..4513)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3842..3863)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4103..4122)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4450..4471
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4514..4909
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5067..5188
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5104..5123)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5158..5177
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5237..5253)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5237..5253)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5250..5272)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5261..5277
                     /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(5285..5291,5292..5309,5310..5315))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5330..5351
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5468..5485)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5639..6227)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5719..5738)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6398..7189,7190..7258))
                     /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     7021..7040
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7259..7363)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7438..7457)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7629..8008
                     /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 accatggcct gtctgatggc cgctttctcg gtcggcaccg
      361 ccatgaatgc cagcagttac tctgcagaga tgacggagcc caagtcggtg tgtgtctcgg
      421 tggatgaggt ggtgtccagc aacatggagg ccactgagac ggacctgctg aatggacatc
      481 tgaaaaaagt agataataac ctcacggaag cccagcgctt ctcctccttg cctcggaggg
      541 cagctgtgaa cattgaattc agggaccttt cctattcggt tcctgaagga ccctggtgga
      601 ggaagaaagg atacaagacc ctcctgaaag gaatttccgg gaagttcaat agtggtgagt
      661 tggtggccat tatgggtcct tccggggccg ggaagtccac gctgatgaac atcctggctg
      721 gatacaggga gacgggcatg aagggggccg tcctcatcaa cggcctgccc cgggacctgc
      781 gctgcttccg gaaggtgtcc tgctacatca tgcaggatga catgctgctg ccgcatctca
      841 ctgtgcagga ggccatgatg gtgtcggcac atctgaagct tcaggagaag gatgaaggca
      901 gaagggaaat ggtcaaggag atactgacag cgctgggctt gctgtcttgc gccaacacgc
      961 ggaccgggag cctgtcaggt ggtcagcgca agcgcctggc catcgcgctg gagctggtga
     1021 acaaccctcc agtcatgttc ttcgatgagc ccaccagcgg cctggacagc gcctcctgct
     1081 tccaggtggt ctcgctgatg aaagggctcg ctcaaggggg tcgctccatc atttgcacca
     1141 tccaccagcc cagcgccaaa ctcttcgagc tgttcgacca gctttacgtc ctgagtcaag
     1201 gacaatgtgt gtaccgggga aaagtctgca atcttgtgcc atatttgagg gatttgggtc
     1261 tgaactgccc aacctaccac aacccagcag attttgtcat ggaggttgca tccggcgagt
     1321 acggtgatca gaacagtcgg ctggtgagag cggttcggga gggcatgtgt gactcagacc
     1381 acaagagaga cctcgggggt gatgccgagg tgaacccttt tctttggcac cggccctctg
     1441 aagaggtaaa gcagacaaaa cgattaaagg ggttgagaaa ggactcctcg tccatggaag
     1501 gctgccacag cttctctgcc agctgcctca cgcagttctg catcctcttc aagaggacct
     1561 tcctcagcat catgagggac tcggtcctga cacacctgcg catcacctcg cacattggga
     1621 tcggcctcct cattggcctg ctgtacttgg ggatcgggaa cgaagccaag aaggtcttga
     1681 gcaactccgg cttcctcttc ttctccatgc tgttcctcat gttcgcggcc ctcatgccta
     1741 ctgttctgac atttcccctg gagatgggag tctttcttcg ggaacacctg aactactggt
     1801 acagcctgaa ggcctactac ctggccaaga ccatggcaga cgtgcccttt cagatcatgt
     1861 tcccagtggc ctactgcagc atcgtgtact ggatgacgtc gcagccgtcc gacgccgtgc
     1921 gctttgtgct gtttgccgcg ctgggcacca tgacctccct ggtggcacag tccctgggcc
     1981 tgctgatcgg agccgcctcc acgtccctgc aggtggccac tttcgtgggc ccagtgacag
     2041 ccatcccggt gctcctgttc tcggggttct tcgtcagctt cgacaccatc cccacgtacc
     2101 tacagtggat gtcctacatc tcctatgtca ggtatgggtt cgaaggggtc atcctctcca
     2161 tctatggctt agaccgggaa gatctgcact gtgacatcga cgagacgtgc cacttccaga
     2221 agtcggaggc catcctgcgg gagctggacg tggaaaatgc caagctgtac ctggacttca
     2281 tcgtactcgg gattttcttc atctccctcc gcctcattgc ctattttgtc ctcaggtaca
     2341 aaatccgggc agagaggtag aacccagctt tcttgtacaa agtggttgat ctagagggcc
     2401 cgcggttcga aggtaagcct atccctaacc ctctcctcgg tctcgattct acgcgtaccg
     2461 gttagtaatg agtttaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa
     2521 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt
     2581 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     2641 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     2701 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg
     2761 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     2821 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     2881 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2941 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3001 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3061 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3121 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3181 aaatgagctg atttaacaaa aatttaacgc gaattatcct cgagctaccg ggtaggggag
     3241 gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc
     3301 tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct
     3361 ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc
     3421 cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt
     3481 ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc
     3541 caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg
     3601 ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg
     3661 cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgact
     3721 ctagacacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     3781 ggtgaggaac taaaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     3841 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     3901 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     3961 cgtgccctgg cccaccctcg tgaccacctt cacctacggc gtgcagtgct tcgcccgcta
     4021 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     4081 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     4141 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     4201 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaaggtct atatcaccgc
     4261 cgacaagcag aagaacggca tcaaggtgaa cttcaagacc cgccacaaca tcgaggacgg
     4321 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     4381 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     4441 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     4501 cgagctgtac aaggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac
     4561 ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc
     4621 tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc
     4681 agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt
     4741 cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct
     4801 tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc
     4861 agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag cacttcgtgg
     4921 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
     4981 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5041 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     5101 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5161 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5221 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5281 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5341 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5401 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5461 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5521 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5581 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5641 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5701 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5761 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5821 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5881 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5941 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6001 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6061 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6121 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6181 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6241 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6301 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6361 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6421 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6481 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6541 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6601 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6661 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6721 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag cttcggttcc caacgatcaa
     6781 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6841 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6901 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6961 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7021 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7081 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7141 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7201 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7261 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7321 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7381 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     7441 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     7501 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     7561 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     7621 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     7681 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     7741 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     7801 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     7861 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     7921 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     7981 ctacgtatta gtcatcgcta ttaccatg
//
