LOCUS       sequence-360362-        6893 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses FLAG tagged FZD4 with linker domain deleted.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6893)
  TITLE     HOLD - Choi lab 2022
REFERENCE   2  (bases 1 to 6893)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pcDNA FZD4-del-linker
FEATURES             Location/Qualifiers
     source          1..6893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             366..395
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             411..428
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(451..468)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    457..681
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      727..1155
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(814..833)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1024..1045
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1164..1184)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1169..1498
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1349..1484
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1411..1430
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1565..2359
                     /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(1619..1638)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2229..2248
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2535..2656
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2572..2591)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2626..2645
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2705..2721)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2705..2721)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2718..2740)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2729..2745
                     /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(2753..2759,2760..2777,2778..2783))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2798..2819
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2936..2953)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3107..3695)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3187..3206)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3866..4657,4658..4726))
                     /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     4489..4508
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4727..4831)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4906..4925)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5097..5476
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5477..5680
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5631..5651
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5725..5744
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5725..5743
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5777..5821
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             5822..5845
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 ggacaaagac agacaagttg gaaaggctaa tggtcaagat cggggtcttc tcagtactgt
       61 acacggttcc tgcaacctgt gtgattgcct gttatttcta tgaaatctca aactgggcac
      121 tctttcgata ttctgcagat gactcaaaca tggcagttga aatgttgaaa atttttatgt
      181 ctttgctcgt gggcatcact tcaggcatgt ggatttggtc tgccaaaact cttcacacgt
      241 ggcaaaagtg ttctaaccga ttggtgaatt ctgggaaggt aaagagagag aagaggggga
      301 atggttgggt gaagccagga aaaggcaacg agactgtggt atagctcgag tctagagggc
      361 ccttcgaaca aaaactcatc tcagaagagg atctgaatat gcataccggt catcatcacc
      421 atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
      481 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
      541 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
      601 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
      661 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
      721 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
      781 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      841 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      901 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      961 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     1021 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     1081 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     1141 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     1201 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     1261 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     1321 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     1381 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     1441 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     1501 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     1561 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     1621 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     1681 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     1741 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     1801 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     1861 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     1921 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     1981 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     2041 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     2101 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     2161 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     2221 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     2281 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     2341 tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc
     2401 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     2461 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     2521 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2581 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2641 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     2701 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     2761 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     2821 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     2881 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2941 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3001 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3061 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3121 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3181 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3241 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3301 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3361 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3421 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3481 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3541 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3601 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3661 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3721 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3781 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3841 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3901 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3961 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4021 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4081 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4141 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4201 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4261 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4321 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4381 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4441 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4501 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4561 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4621 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4681 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4741 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4801 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4861 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     4921 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     4981 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     5041 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
     5101 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     5161 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     5221 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     5281 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     5341 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     5401 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     5461 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     5521 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     5581 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     5641 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
     5701 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
     5761 gttaagcttg gtaccaatgt ctgcacttct gatcctagct cttgttggag ctgcagttgc
     5821 tgactacaaa gacgatgacg acaagcttgc ggccgcagag gagcggcgct gcgaccccat
     5881 ccgcatcgcc atgtgccaga acctcggcta caacgtgacc aagatgccca acttagtggg
     5941 acacgagctg cagacagacg ccgagctgca gctgacaact ttcacgccgc tcatccagta
     6001 cggctgctcc agccagctgc agttcttcct ttgttcggtt tatgtgccaa tgtgcacaga
     6061 gaagatcaac atccccatcg gcccgtgcgg tggcatgtgc ctttcagtca agagacgctg
     6121 tgaaccagtc ctgagagaat ttgggtttgc ctggcccgac accctgaact gcagcaagtt
     6181 cccgccccag aacgaccaca accacatgtg catggaagga ggttcaggag gttgtggcta
     6241 cgatgctggc ttgtacagcc gctcagctaa ggagttcacg gatatttgga tggctgtgtg
     6301 ggccagcctc tgcttcatct ccaccacctt caccgtgctg accttcctga ttgattcatc
     6361 caggttttct taccctgagc gccccatcat atttctcagt atgtgctata atatttatag
     6421 cattgcttat attgttcggc tgactgtagg ccgggaaagg atatcctgtg attttgaaga
     6481 ggcggcagag cccgttctca tccaagaagg acttaagaac acaggatgtg caataatttt
     6541 cttgctgatg tacttttttg gaatggccag ctccatttgg tgggttattc tgacactcac
     6601 ttggtttttg gcagccggac tcaagtgggg tcatgaagcc attgaaatgc acagttctta
     6661 tttccacatc gcagcctggg ctattcccgc agtgaaaacc attgtcatct tgattatgag
     6721 actagtggat gccgatgaac tgactggctt gtgctatgtt gggaaccaaa acctagatgc
     6781 cctcactggc tttgtggtgg ctcctctctt tacgtatttg gtgattggaa cgctgttcat
     6841 tgcggcgggt ttggtggcct tattcaaaat tcggtccaat cttcaaaaag acg
//
