LOCUS       sequence-349724-        6971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector encoding JAM2 and EmGFP-Bsd.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6971)
  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 6971)
  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: JAM2_pcDNA6.2/EmGFP-Bsd
FEATURES             Location/Qualifiers
     source          1..6971
                     /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(1313..1337)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1363..1404
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1444..1463)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1488..1536
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1738..2166
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1825..1844)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2035..2056
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2181..2680
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2216..2239)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2613..2632
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        2694..2741
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(2760..2762,2763..2765,2766..3476)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2805..2826)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3066..3085)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3413..3434
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3477..3872
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4030..4151
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4067..4086)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4121..4140
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4200..4216)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4200..4216)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4213..4235)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4224..4240
                     /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(4248..4254,4255..4272,4273..4278))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4293..4314
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4431..4448)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4602..5190)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4682..4701)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5361..6152,6153..6221))
                     /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     5984..6003
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6222..6326)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6401..6420)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6592..6971
                     /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 tccgcggccg cccccttcac ccctcctcag agcagccggc
      361 tgccgccccg ggaagatggc gaggaggagc cgccaccgcc tcctcctgct gctgctgcgc
      421 tacctggtgg tcgccctggg ctatcataag gcctatgggt tttctgcccc aaaagaccaa
      481 caagtagtca cagcagtaga gtaccaagag gctattttag cctgcaaaac cccaaagaag
      541 actgtttcct ccagattaga gtggaagaaa ctgggtcgga gtgtctcctt tgtctactat
      601 caacagactc ttcaaggtga ttttaaaaat cgagctgaga tgatagattt caatatccgg
      661 atcaaaaatg tgacaagaag tgatgcgggg aaatatcgtt gtgaagttag tgccccatct
      721 gagcaaggcc aaaacctgga agaggataca gtcactctgg aagtattagt ggctccagca
      781 gttccatcat gtgaagtacc ctcttctgct ctgagtggaa ctgtggtaga gctacgatgt
      841 caagacaaag aagggaatcc agctcctgaa tacacatggt ttaaggatgg catccgtttg
      901 ctagaaaatc ccagacttgg ctcccaaagc accaacagct catacacaat gaatacaaaa
      961 actggaactc tgcaatttaa tactgtttcc aaactggaca ctggagaata ttcctgtgaa
     1021 gcccgcaatt ctgttggata tcgcaggtgt cctgggaaac gaatgcaagt agatgatctc
     1081 aacataagtg gcatcatagc agccgtagta gttgtggcct tagtgatttc cgtttgtggc
     1141 cttggtgtat gctatgctca gaggaaaggc tacttttcaa aagaaacctc cttccagaag
     1201 agtaattctt catctaaagc cacgacaatg agtgaaaatg atttcaagca cacaaaatcc
     1261 tttataattt aaagactcca ctttagagat acaccaaagg gtgggcgcgc cgacccagct
     1321 ttcttgtaca aagtggttga tctagagggc ccgcggttcg aaggtaagcc tatccctaac
     1381 cctctcctcg gtctcgattc tacgcgtacc ggttagtaat gagtttaaac gggggaggct
     1441 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     1501 acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     1561 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     1621 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     1681 gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg
     1741 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1801 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     1861 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1921 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     1981 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2041 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     2101 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2161 cgaattatcc tcgagctacc gggtagggga ggcgcttttc ccaaggcagt ctggagcatg
     2221 cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc ctcgcacaca
     2281 ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc
     2341 ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac
     2401 gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca ccgctgagca
     2461 atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg
     2521 ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg
     2581 ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc
     2641 tgccgcgctg ttctcctctt cctcatctcc gggcctttcg actctagaca cgtgttgaca
     2701 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca
     2761 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     2821 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     2881 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2941 tcgtgaccac cttcacctac ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc
     3001 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     3061 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     3121 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     3181 agctggagta caactacaac agccacaagg tctatatcac cgccgacaag cagaagaacg
     3241 gcatcaaggt gaacttcaag acccgccaca acatcgagga cggcagcgtg cagctcgccg
     3301 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     3361 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     3421 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaaggcca
     3481 agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca
     3541 tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct
     3601 tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg
     3661 gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga
     3721 acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg
     3781 ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat
     3841 tgctgccctc tggttatgtg tgggagggct aagcacttcg tggccgagga gcaggactga
     3901 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3961 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4021 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4081 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4141 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     4201 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4261 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4321 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4381 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4441 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4501 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4561 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4621 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4681 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4741 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4801 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4861 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4921 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4981 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5041 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5101 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5161 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5221 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5281 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5341 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5401 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5461 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5521 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5581 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5641 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5701 tcgtttggta tggcttcatt cagcttcggt tcccaacgat caaggcgagt tacatgatcc
     5761 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5821 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5881 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5941 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6001 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6061 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6121 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6181 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6241 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6301 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     6361 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     6421 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     6481 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     6541 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
     6601 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     6661 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     6721 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     6781 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     6841 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     6901 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     6961 ctattaccat g
//
