LOCUS       sequence-360389-        7589 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses FLAG tagged FZD6 swapped with FZD4 CRDlinker domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7589)
  TITLE     HOLD - Choi lab 2022
REFERENCE   2  (bases 1 to 7589)
  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 FZD6_4CRDlinker
FEATURES             Location/Qualifiers
     source          1..7589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(24..44)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        29..358
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      209..344
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     271..290
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             425..1219
                     /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(479..498)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1089..1108
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1395..1516
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1432..1451)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1486..1505
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1565..1581)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1565..1581)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1578..1600)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1589..1605
                     /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(1613..1619,1620..1637,1638..1643))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1658..1679
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1796..1813)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1967..2555)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2047..2066)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2726..3517,3518..3586))
                     /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     3349..3368
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3587..3691)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3766..3785)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3957..4336
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4337..4540
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4491..4511
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4585..4604
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4585..4603
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4636..4680
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             4681..4704
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6860..6877
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6900..6917)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6906..7130
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(7176..7589,1..15)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7263..7282)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7473..7494
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
       61 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
      121 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      181 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
      241 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
      301 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
      361 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
      421 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
      481 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
      541 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
      601 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
      661 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
      721 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
      781 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
      841 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
      901 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
      961 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     1021 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     1081 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     1141 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     1201 tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc
     1261 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     1321 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     1381 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     1441 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     1501 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     1561 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     1621 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     1681 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     1741 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     1801 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1861 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1921 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1981 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2041 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2101 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2161 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2221 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2281 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2341 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2401 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2461 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2521 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2581 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2641 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2701 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2761 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     2821 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     2881 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     2941 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3001 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3061 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3121 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3181 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3241 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     3301 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     3361 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     3421 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     3481 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     3541 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     3601 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3661 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     3721 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     3781 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     3841 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     3901 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
     3961 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     4021 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     4081 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     4141 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     4201 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     4261 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     4321 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     4381 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     4441 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     4501 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
     4561 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
     4621 gttaagcttg gtaccatgtc tgcacttctg atcctagctc ttgttggagc tgcagttgct
     4681 gactacaaag acgatgacga caagcttgcg gccgcagagg agcggcgctg cgaccccatc
     4741 cgcatcgcca tgtgccagaa cctcggctac aacgtgacca agatgcccaa cttagtggga
     4801 cacgagctgc agacagacgc cgagctgcag ctgacaactt tcacgccgct catccagtac
     4861 ggctgctcca gccagctgca gttcttcctt tgttcggttt atgtgccaat gtgcacagag
     4921 aagatcaaca tccccatcgg cccgtgcggt ggcatgtgcc tttcagtcaa gagacgctgt
     4981 gaaccagtcc tgagagaatt tgggtttgcc tggcccgaca ccctgaactg cagcaagttc
     5041 ccgccccaga acgaccacaa ccacatgtgc atggaaggac caggtgatga agaggttccc
     5101 ttgccccaca agactcccat ccagcccggg gaagagtgcc actccgtggg aagcaattct
     5161 gatcagtaca tctgggtgaa gaggagcctg aactgtgttc tcaagtgccc caacatgtat
     5221 tttaaaagtg atgagctaga gtttgcaaaa agttttattg gaacagtttc aatattttgt
     5281 ctttgtgcaa ctctgttcac attccttact tttttaattg atgttagaag attcagatac
     5341 ccagagagac caattatata ttactctgtc tgttacagca ttgtatctct tatgtacttc
     5401 attggatttt tgctgggcga tagcacagcc tgcaataagg cagatgagaa gctagaactt
     5461 ggtgacactg ttgtcctagg ctctcaaaat aaggcttgca ccgttttgtt catgcttttg
     5521 tattttttca caatggctgg cactgtgtgg tgggtgattc ttaccattac ttggttctta
     5581 gctgcaggaa gaaaatggag ttgtgaagcc atcgagcaaa aagcagtgtg gtttcatgct
     5641 gttgcatggg gaacaccagg tttcctgact gttatgcttc ttgctctgaa caaagttgaa
     5701 ggagacaaca ttagtggagt ttgctttgtt ggcctttatg acctggatgc ttctcgctac
     5761 tttgtactct tgccactgtg cctttgtgtg tttgttgggc tctctcttct tttagctggc
     5821 attatttcct taaatcatgt tcgacaagtc atacaacatg atggccggaa ccaagaaaaa
     5881 ctaaagaaat ttatgattcg aattggagtc ttcagcggct tgtatcttgt gccattagtg
     5941 acacttctcg gatgttacgt ctatgagcaa gtgaacagga ttacctggga gataacttgg
     6001 gtctctgatc attgtcgtca gtaccatatc ccatgtcctt atcaggcaaa agcaaaagct
     6061 cgaccagaat tggctttatt tatgataaaa tacctgatga cattaattgt tggcatctct
     6121 gctgtcttct gggttggaag caaaaagaca tgcacagaat gggctgggtt ttttaaacga
     6181 aatcgcaaga gagatccaat cagtgaaagt cgaagagtac tacaggaatc atgtgagttt
     6241 ttcttaaagc acaattctaa agttaaacac aaaaagaagc actataaacc aagttcacac
     6301 aagctgaagg tcatttccaa atccatggga accagcacag gagctacagc aaatcatggc
     6361 acttctgcag tagcaattac tagccatgat tacctaggac aagaaacttt gacagaaatc
     6421 caaacctcac cagaaacatc aatgagagag gtgaaagcgg acggagctag cacccccagg
     6481 ttaagagaac aggactgtgg tgaacctgcc tcgccagcag catccatctc cagactttct
     6541 ggggaacagg tcgacgggaa gggccaggca ggcagtgtat ctgaaagtgc gcggagtgaa
     6601 ggaaggatta gtccaaagag tgatattact gacactggcc tggcacagag caacaatttg
     6661 caggtcccca gttcttcaga accaagcagc ctcaaaggtt ccacatctct gcttgttcac
     6721 ccagtttcag gagtgagaaa agagcaggga ggtggttgtc attcagatac ttgagaattc
     6781 tgggaaggta aagagagaga agagggggaa tggttgggtg aagccaggaa aaggcaacga
     6841 gactgtggta tagctcgagc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     6901 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     6961 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     7021 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     7081 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     7141 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     7201 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     7261 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     7321 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     7381 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     7441 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     7501 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     7561 ttggttaaaa aatgagctga tttaacaaa
//
