LOCUS       sequence-351439-        7976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian lentiviral expression vector encoding CRYZL1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7976)
  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 7976)
  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: CRYZL1_pLENTI-CAG-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..7976
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          145..1161
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1085..1107
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1169..1188
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1277..1301
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2355..2379)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2399..2422
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2457..3006
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2601..2618)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2828..2847
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3009..3011,3012..3014,3015..3728)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3009..3011,3012..3014,3015..3725)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3054..3075)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3315..3334)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3662..3683
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3805..4393
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3858..3878)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4276..4287)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4396..4412)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4397..4413)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             4602..4835
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4907..5028
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4944..4963)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4998..5017
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5068..5203
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5223..5242)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5224..5242)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(5265..5623)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5305..5326)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5517..5536
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5650..5754
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5755..5823,5824..6615)
                     /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     complement(5973..5992)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6786..7374
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7275..7294
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7549..7852
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat
       61 cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat
      121 aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc
      181 tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg
      241 agcgggcggg acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt
      301 gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg
      361 gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc
      421 gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt
      481 gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt gcgggggggg ctgcgagggg
      541 aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg
      601 gtcgggctgc aaccccccct gcacccccct ccccgagttg ctgagcacgg cccggcttcg
      661 ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg
      721 caggtggggg tgccgggcgg ggcggggccg cctcgggccg gggagggctc gggggagggg
      781 cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt
      841 ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg tgcggagccg
      901 aaatctggga ggcgccgccg caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc
      961 ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc
     1021 cctctccagc ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg
     1081 gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat
     1141 gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat
     1201 tttggcaaag aattccccta gccgcggaat ctagactgcc ggatcgaatt gtcgacgaat
     1261 tcggatctgg ccaatcacaa gtttgtacaa aaaagcaggc accatgaaag gcttatattt
     1321 ccaacagagt tccacagatg aagaaataac atttgtattt caagaaaagg aagatcttcc
     1381 tgttacagag gataactttg tgaaacttca agttaaagct tgtgctctga gccagataaa
     1441 tacaaagctt ctggcagaaa tgaagatgaa aaaggattta tttcctgttg ggagagaaat
     1501 tgctggaatt gtattagatg ttggaagcaa ggtatcattc tttcaaccag atgatgaagt
     1561 agttggaatt ttgcccctgg actctgaaga ccctggactt tgtgaagttg ttagagtaca
     1621 tgagcattac ttggttcata aaccagaaaa ggtcacatgg acggaagcag caggaagcat
     1681 tcgggatgga gtgcgtgcct atacagctct gcattatctt tctcatctct ctcctggaaa
     1741 atcagtgctg ataatggatg gagcaagtgc atttggtaca atagctattc agttagcaca
     1801 tcatagagga gccaaagtga tttcaacagc atgcagcctt gaagataagc agtgccttga
     1861 aagattcaga cctcccatag cccgagtgat tgatgtatct aatgggaaag ttcatgttgc
     1921 tgaaagctgt ttggaagaaa caggtggcct gggagtagat attgtcctag atgctggagt
     1981 gagattatat agtaaagatg atgaaccagc tgtaaaacta caactactac cacataaaca
     2041 tgatatcatc acacttcttg gtgttggagg ccactgggta acaacagaag aaaaccttca
     2101 gttggatcct ccagatagcc actgcctttt cctcaaggga gcaacgttag ctttcctgaa
     2161 tgatgaagtt tggaatttgt caaatgtaca acagggaaaa tatctttgta tcttaaagga
     2221 tgtgatggag aagttatcaa ctggtgtttt cagacctcag ttggatgaac ccattccact
     2281 gtatgaggca aaagtttcca tggaagctgt tcagaaaaat caaggaagaa aaaagcaagt
     2341 tgttcaattt tagaacccag ctttcttgta caaagtggtg ggaggaggtg gatctggcga
     2401 ctacaaggac gacgatgaca agtgactcga gcgggatcaa ttccgccccc cccctaacgt
     2461 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac
     2521 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
     2581 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa
     2641 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
     2701 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
     2761 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
     2821 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc
     2881 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag
     2941 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     3001 ataataccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     3061 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     3121 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     3181 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     3241 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     3301 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     3361 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     3421 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     3481 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     3541 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     3601 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     3661 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     3721 acaagtaaag cggccgcgac tctagagtcg acctgcaggc atgcaagctt caggtagccg
     3781 gctaacgtta tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     3841 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3901 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     3961 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     4021 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     4081 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     4141 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     4201 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     4261 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     4321 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     4381 gccgcctccc cgcatcgata ccgtcgacct cgatcgagac ctagaaaaac atggagcaat
     4441 cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga
     4501 ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc
     4561 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     4621 ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
     4681 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     4741 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     4801 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
     4861 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
     4921 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4981 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
     5041 ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     5101 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     5161 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt
     5221 cgccctatag tgagtcgtat tagcatgtga gcaaaaggcc agcataaatc aaaagaatag
     5281 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     5341 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     5401 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     5461 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     5521 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     5581 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     5641 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5701 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5761 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5821 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5881 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5941 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     6001 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6061 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6121 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6181 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6241 tgggaaccga agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     6301 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6361 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6421 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6481 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6541 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6601 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6661 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6721 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6781 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6841 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6901 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6961 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7021 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7081 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7141 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7201 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7261 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     7321 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7381 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     7441 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata
     7501 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     7561 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     7621 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     7681 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     7741 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     7801 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt
     7861 gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta
     7921 tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggg
//
