LOCUS       sequence-224019-        7026 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human FZD4  in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7026)
  AUTHORS   Lai MB, Zhang C, Shi J, Johnson V, Khandan L, McVey J, Klymkowsky 
            MW, Chen Z, Junge HJ
  TITLE     TSPAN12 Is a Norrin Co-receptor that Amplifies Frizzled4 Ligand 
            Selectivity and Signaling.
  JOURNAL   Cell Rep. 2017 Jun 27;19(13):2809-2822. doi: 
            10.1016/j.celrep.2017.06.004.
  PUBMED    28658627
REFERENCE   2  (bases 1 to 7026)
  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"; doi: 
            "10.1016/j.celrep.2017.06"; journalName: "Cell Rep"; date: 
            "2017-06-27- 27"; volume: "19"; issue: "13"; pages: "2809-2822"
COMMENT     Sequence Label: pcDNA 3.3 hFZD4
FEATURES             Location/Qualifiers
     source          1..7026
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        50..429
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        430..633
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     584..604
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     630..654
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(2412..2431)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2456..2504
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2706..3134
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2793..2812)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3003..3024
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3143..3163)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3148..3477
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3328..3463
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3390..3409
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3544..4338
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3598..3617)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4208..4227
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4514..4635
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4551..4570)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4605..4624
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4684..4700)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4684..4700)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4697..4719)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4708..4724
                     /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(4732..4738,4739..4756,4757..4762))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4777..4798
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4915..4932)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5086..5674)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5166..5185)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5845..6636,6637..6705))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6468..6487
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6706..6810)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6885..6904)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagg atcgaaccct tccaccatgg cctggcgggg cgcagggccg agcgtcccgg
      781 gggcgcccgg gggcgtcggt ctcagtctgg ggttgctcct gcagttgctg ctgctcctgg
      841 ggccggcgcg gggcttcggg gacgaggaag agcggcgctg cgaccccatc cgcatctcca
      901 tgtgccagaa cctcggctac aacgtgacca agatgcccaa cctggttggg cacgagctgc
      961 agacggacgc cgagctgcag ctgacaactt tcacaccgct catccagtac ggctgctcca
     1021 gccagctgca gttcttcctt tgttctgttt atgtgccaat gtgcacagag aagatcaaca
     1081 tccccattgg cccatgcggc ggcatgtgtc tttcagtcaa gagacgctgt gaacccgtcc
     1141 tgaaggaatt tggatttgcc tggccagaga gtctgaactg cagcaaattc ccaccacaga
     1201 acgaccacaa ccacatgtgc atggaagggc caggtgatga agaggtgccc ttacctcaca
     1261 aaacccccat ccagcctggg gaagagtgtc actctgtggg aaccaattct gatcagtaca
     1321 tctgggtgaa aaggagcctg aactgtgtgc tcaagtgtgg ctatgatgct ggcttataca
     1381 gccgctcagc caaggagttc actgatatct ggatggctgt gtgggccagc ctgtgtttca
     1441 tctccactgc cttcacagta ctgaccttcc tgatcgattc ttctaggttt tcctaccctg
     1501 agcgccccat catatttctc agtatgtgct ataatattta tagcattgct tatattgtca
     1561 ggctgactgt aggccgggaa aggatatcct gtgattttga agaggcagca gaacctgttc
     1621 tcatccaaga aggacttaag aacacaggat gtgcaataat tttcttgctg atgtactttt
     1681 ttggaatggc cagctccatt tggtgggtta ttctgacact cacttggttt ttggcagcag
     1741 gactcaaatg gggtcatgaa gccattgaaa tgcacagctc ttatttccac attgcagcct
     1801 gggccatccc cgcagtgaaa accattgtca tcttgattat gagactggtg gatgcagatg
     1861 aactgactgg cttgtgctat gttggaaacc aaaatctcga tgccctcacc gggttcgtgg
     1921 tggctcccct ctttacttat ttggtcattg gaactttgtt cattgctgca ggtttggtgg
     1981 ccttgttcaa aattcggtca aatcttcaaa aggatgggac aaagacagac aagttagaaa
     2041 gactgatggt caagattggg gtgttctcag tactgtacac agttcctgca acgtgtgtga
     2101 ttgcctgtta tttttatgaa atctccaact gggcactttt tcggtattct gcagatgatt
     2161 ccaacatggc tgttgaaatg ttgaaaattt ttatgtcttt gttggtgggc atcacttcag
     2221 gcatgtggat ttggtctgcc aaaactcttc acacgtggca gaagtgttcc aacagattgg
     2281 tgaattctgg aaaggtaaag agagagaaga gaggaaatgg ttgggtgaag cctggaaaag
     2341 gcagtgagac tgtggtataa aagggttcga tccctaccgg ttagtaatga gtttaaacgg
     2401 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     2461 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     2521 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     2581 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     2641 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     2701 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2761 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     2821 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     2881 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     2941 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3001 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3061 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3121 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3181 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3241 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3301 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3361 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     3421 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     3481 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     3541 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3601 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3661 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3721 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3781 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3841 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3901 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3961 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4021 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4081 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4141 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4201 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4261 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4321 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     4381 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4441 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4501 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4561 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4621 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     4681 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4741 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4801 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4861 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4921 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4981 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5041 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5101 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5161 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5221 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5281 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5341 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5401 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5461 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5521 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5581 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5641 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5701 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5761 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5821 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5881 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5941 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6001 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6061 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6121 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6181 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6241 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6301 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6361 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6421 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6481 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6541 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6601 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6661 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6721 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6781 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6841 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
     6901 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     6961 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     7021 cttagg
//
