LOCUS       Exported                9815 bp ds-DNA     circular SYN 30-MAY-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9815)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jun 5, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9815
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    15..275
                     /label=5'Tol2 DNA
                     /note="5'Tol2 DNA"
     misc_feature    15..174
                     /label=5'Tol2 arm
     primer_bind     complement(255..291)
                     /label=pmini Tol2 R
     misc_feature    276..298
                     /label=MCS
                     /note="MCS"
     misc_feature    300..4308
                     /label=4 kb Wnt3 promoter
                     /note="4 kb Wnt3 promoter"
     misc_feature    363..368
                     /label=TCF BS (CATTGT)
     misc_feature    410..415
                     /label=TCF BS (CATTGT)
     misc_feature    464..471
                     /label=SP5 BS (GGGGGTGG)
     misc_feature    495..501
                     /label=TCF BS (AACAAAG)
     misc_feature    complement(580..639)
                     /label=Vec_minitol2(C80A)RP1
                     /note="Vec_minitol2(C80A)RP1"
     misc_feature    complement(580..619)
                     /label=Vec_mtol2(C80)RP1_Rev
                     /note="Vec_mtol2(C80)RP1_Rev"
     misc_feature    600..619
                     /label=VecMiniTol2(C80A)FP2
                     /note="VecMiniTol2(C80A)FP2"
     misc_feature    complement(614..633)
                     /label=Vec_mTol2(C80)RP1_R3
                     /note="Vec_mTol2(C80)RP1_R3"
     misc_feature    746..759
                     /label=PDX1 (GGCTCTAATGAaAa)
     misc_feature    1114..1119
                     /label=TCF BS (CATTGT)
     misc_feature    1374..1387
                     /label=ETF-A(GGCTTTCcTCAaAG)
     misc_feature    1628..1633
                     /label=TCF BS (CATTGT)
     misc_feature    1701..1714
                     
     misc_feature    2049..2054
                     /label=TCF BS (CATTGT)
     misc_feature    2314..2321
                     /label=TCF BS (GATTGGCT)
     misc_feature    2744..2757
                     /label=SMAD3-SMAD4 (CAGAtAGACAGACA)
     misc_feature    2752..2765
                     /label=SMAD3-SMAD4 (CAGACAGACAGgCt)
     misc_feature    2883..2895
                     /label=C/EBP (TTTTGTAATGGGt)
     misc_feature    3362..3369
                     /label=TCF BS (CCTTTGGG)
     misc_feature    3769..3776
                     /label=SP5 BS (GGGGGTGG)
     misc_feature    3793..3798
                     /label=SP5 BS (GGGCAG)
     misc_feature    3848..3855
                     /label=TCF BS (GATTGGCT)
     misc_feature    4308..4313
                     /label=BamH1
                     /note="BamH1"
     misc_feature    4309..4313
                     /label=MCS2
                     /note="MCS2"
     LTR             4314..4335
                     
                     /note="5'UTR Fzd1"
     misc_feature    4336..5949
                     /label=Fzd1 ORF 1614 bp
                     /note="Fz ORF without stop codon"
     misc_feature    5950..5973
                     /label=(GGGS)2 linker
                     /note="(GGGS)2 linker"
     misc_feature    5974..6684
                     /label=mApple
                     /note="mApple"
     misc_feature    complement(6665..6723)
                     /label=Insert_mApple(mTol2)RP
                     /note="Insert_mApple(mTol2)RP"
     misc_feature    6685..6692
                     /label=Not1
                     /note="Not1"
     misc_feature    6693..6889
                     /label=3' Tol2 DNA
                     /note="3' Tol2 DNA"
     promoter        complement(6940..6958)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6976..6992)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7000..7016
                     /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(7024..7054)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(7378..7966)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8137..8997)
                     /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"
     promoter        complement(8998..9102)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(9180..9635)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     9776..9792
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(9799..9815,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg ggcccagagg tgtaaagtac ttgagtaatt ttacttgatt actgtactta
       61 agtattattt ttggggattt ttactttact tgagtacaat taaaaatcaa tacttttact
      121 tttacttaat tacatttttt tagaaaaaaa agtacttttt actccttaca attttattta
      181 cagtcaaaaa gtacttattt tttggagatc acttcattct attttccctt gctattacca
      241 aaccaattga attgcgctga tgcccagttt aatttaaata gatctggcca tctagagcaa
      301 gcttcaataa aagaagtgtc tgttgtgggg ccgagggtcc tggggggctg cggagcttta
      361 ggcattgtcc tggagggtga ttgagagcga gaggagggga gccgagaagc attgtgcgag
      421 gacggagcct ttctaaaggt ggttattagg agagaaatgg tgtgggggtg gctgttgact
      481 cttcatcttg tgtgaacaaa gtatctggcg tatagctttc tctctctctc tctcgctctc
      541 ttgcggcttt cgctttacga cggccatgaa tactctcggc gggctcgccc agacctgcgc
      601 gaaggcttat tgtccggctt aagtgcggtt taattaagcg agctctgtgt cctgagagaa
      661 gagaggagct ttagagcgct cgggtcctgc ggagatgcag aggatgatgc gtaatatcag
      721 ggctattgaa ggggaaagtg tttgcggctc taatgaaaaa ggtttgtttg ggggggatga
      781 gtggggaggg gattgaggag tgtctggcct ttgtctccag cttacaaagt agggacaaaa
      841 gcagtgatat tatacaaatg cacccatttc caggacatga tgcgaggacg ggggcctgga
      901 gatgggtcgg actgctccat ctcagccccg ttggcacctt acaaaatgat tttttccccc
      961 caccacaaag ataggcattg gggataaacc ccgagagctg gggcttctgg gtaccgcaaa
     1021 tttgggggtt gtggacttaa gcggattttc tctgcaactg taagaacaag ctaattcctc
     1081 agacctcctt tatctctaac atggtaataa caaacaatga aattctgaag agaaaggggc
     1141 aaaggaagag gagggaggag agacgcagac acaaaagaag gagagatgga gggaatgaaa
     1201 gagaggagag acgtgtgtga ggagagggtg ggcttgggcg tgttaagaaa ctttctcgct
     1261 ctagtgttat gtgtgaacac tctgctaatc caggttttat tgcttgcctg tgtccggcca
     1321 cactgtccgg attgagtgtg tgtcagggat agtgggggtc tacacatgtc cctctttgag
     1381 gaaagcctga gaaagagccg gacctggtta cggccgtttt ggggtcgcct aagaaacctg
     1441 caccacaggg gtcctctgtg tatgtctctg aattcaacgt gcatgttgct agcaaacaca
     1501 tacggattaa tgctaatgat gaaggtgtgg ctatattagg tacaaaaaaa aaaactttga
     1561 caatagttgt ataaactcct gtcaacagtc catcaatgta tgaagcagaa ctcgctttga
     1621 aaccaaacat tgttgtgtgg gtccatttgg accagtttaa ggaaatcttt tgccctataa
     1681 attccagcct gcatgaattt ttatgattat tgctcataaa catactttta aaaatgtgca
     1741 ttttaactaa ctatcaactt gtgcccgtcc actcaaagga agccaaaagg gtgtctctcc
     1801 cagatcacag ggactacttt gtgggtgtgt agggtgatgt attagctagg cgctaataaa
     1861 atgcaggagt gtccacaaac atgacttaat gagggaggat gagagactcc tgacccagag
     1921 gtcaaagctc cgggcgactc tctttattcc cattgatctg ctggtgcact tcatttccat
     1981 gctctagggc ccacttcagt aaattcatta agtgcgattg atacgagaat caagacttcc
     2041 tccaaaacac aatggctatg ctaggaggga atggtcttgt ttgtgctgca gctatgtcaa
     2101 gcatctctgg cttttgtcct ggtttcgcac cattgcggtg catgctgggg tttgagtcct
     2161 cgagattcta atcgtaagct catttccagc tggttcgttg tgttgcgccc ctatggcctt
     2221 gacctgggaa acgtggtttc agtgttgttg agatgccgac cgcagaccgt tccttgcctc
     2281 cactcgtaaa ttacgctccg atcaggtttt cccagccaat cctggcgagc agatctgcat
     2341 ctgtcactct gcaagaatgc tgaatcaaac cgctgatcta ggcacaatac agctgatgga
     2401 aagcagatct ttattccaac atcaggaacg tagcacgagc cgagtgcttc tggaagctgc
     2461 aatttaacca acaggttcat atctgatgag atcaaggatg catcatcctg gaagaaacac
     2521 tgtgatgaac atttaagatg tcagaatgac ccgtcatgcc tctgagctga gtccccgcac
     2581 agcgcttatt cttttgaaga gggagaataa aagcaaaaaa acagaggcga tggaaatatg
     2641 acatccagac cgcggtcaga actgtgagtg tctctcctgg gctcctatgg ttgtggtgca
     2701 cagtccatgt ccctgctatg agttagatag acagatgcac tggcagatag acagacagac
     2761 aggctttact acttggtcgc tggattgcag gaaagagctt tatccaacca gagtcaagca
     2821 agcaacatgg gacaataaca taattaaata ttctggggaa aagtggcact tatgcaaatg
     2881 ttttttgtaa tgggttggag aaaattggcc ggaacctaca taatttttaa tgacaggatg
     2941 attcatttgt gaaattgtaa ttcaaagaaa ttcagtaaac taactaacaa tctaaaaaat
     3001 tctttgattt gcagtttaat tctacttcct aacattgatt tgtttcaact ttattcaaat
     3061 ggcaaaacta aattagtact caaagctcca aatgatatga ttcctagact gtttaaagtg
     3121 ctaaaagtgc caaaacttac agctacccac atgaacttat atttcattct gcctccattt
     3181 gcaattcctc cagtgtttgt acctgaaact gttgtcagtt ctcacctctg tcctgccgtt
     3241 cattctgaca cagagactct atatttgaat gtattgtcaa cttctaatgc cgtgtctact
     3301 actctgtgct gtctgctctc atcccatccg aatgacaata gtgtttatgt aaaatgtgct
     3361 gcctttgggc cgctggggca cagaacactg catgcctggc cctgccacac caccttttac
     3421 ggctttttat tgtgttgcct ttgtgtcaca aggggaggac gcggccatca aaattctcca
     3481 agacaaaatt cttgaatgta tgtgcacgca cacgcatccg gaacaattag ttcaggagca
     3541 cgggtgctta aaaaacggag acgacacttc tcgccaccac ctcctcgccg agcagcaggc
     3601 ccgagtctgg cacagctggg tttccttgat gctaacgcag acaatataga tcttaataag
     3661 acaaatctac ctctccgaaa ctcagtcctg acaccctttc attcttattc cctcccagcc
     3721 tcctctctct ctctctctct ctttctttct ttctttgcta cattaccagg gggtggtgca
     3781 tgattgacag atgggcagga ataataattt tttttttttt tttgcgtaac ctaaggccat
     3841 gctctgtgat tggcttgcca gctgacatct ccaaacctct cttgtcgttt ttttcccctc
     3901 tctctctctc gctttctctc tctcactctc tctctctaat attggcsctg atgatcctaa
     3961 ggggaaaaaa gtggggagag aaaaagatat gtaccttagg gaagaacctc agtaagaggg
     4021 agggaatcag aggacgagta ggagcgtgtg tgcgagtgtg tgaaagagac acttactctc
     4081 ctaactacta ctggaagaca cacacttttc tcacccaact caaagcagca gcaaaccttg
     4141 gagaattgtt ttatttttgt tttttcgcga accccccyaa aaaaaattcc taattattct
     4201 ctcaagcatc catctttgcc ttgccgggat ttaattttca ccccctctct aggaatccta
     4261 tgtactgatc aatagaccct ttccatccac acaagacata cctccgcgga tcctttctca
     4321 ccgcacaccg aaagtatggc cgcgcgcgcg ctcttcctga tcgccctcgg agctctgagc
     4381 gcccgcgcgc agtacggaga ccgcggaatg tcggtgcccg agcatggctt ctgccagccc
     4441 atctccatcc ctctgtgcac ggacatcgcc tacaacgaga ccatcatgcc caacctgctc
     4501 ggccacacca accaggagga cgcgggactg gaagtgcacc agttttaccc gctggtgaaa
     4561 gtgcagtgct cgccagacct caagttcttc ctgtgctcca tgtacgcgcc ggtgtgcacg
     4621 gtgctggagc aggcgctgcc gccgtgccgg tccctctgcg agcgcgcgcg tcagggctgc
     4681 gaggcgctga tgaataaatt cggcttccag tggccggaga gcctggcgtg cgagtccttc
     4741 ccggtgcacg gcggagagct ctgcgtggga cagaacacca ccgggccgag ccccccagtc
     4801 agcccgactc ccgacgcgac gcaggcgcct ccggagccct cccagcgcgg ccgcttccag
     4861 tgcccgccgt cgctcagagt ccccccgtac ctcaactacc gcttcatggg agaggacaac
     4921 tgcggagcgc cctgcgagcc cagaaaactg cacggcatga tgtacttcag cgaggacgag
     4981 cagaagtttg cgcgcatctg gatcgggatc tggtccgtcc tatgctgcgc ttcaacttta
     5041 ttcacagttc tgacttatct ggtggacatg aagcgcttca gctacccgga gagacccatc
     5101 atattcctgt ccggctgcta caccatggtg tccatcgcct acatcgccgg atttctcctg
     5161 gaggacaagg ttgtctgcaa cgagcagttc gagaacgagt tcaggactgt ggttcagggc
     5221 accaagaagg aaggctgcac catattgttc atgatgctgt acttcttcag catgtccagc
     5281 tccatctggt gggtgatcct ggcgctcacc tggttcctgg cggccgggat gaaatggggt
     5341 cacgaggcca tcgaggccaa ctcgcagtac ttccacctcg cggcctgggc agtgcccgcc
     5401 atcaaaacca tcaccatcct ggctgtgggt caggtggacg gggacgtgct cagcggggtg
     5461 tgctttgtgg gcatcaacag cgtggacgcg ctgcgcggat tcgtgctcgc gccgctcttc
     5521 gtgtacctgt tcatcggcac ctctttcctc ctggcgggat tcgtgtcgtt gttccgcatc
     5581 aggaccatca tgaagcacga cggcactaaa acggagaagc tggagaagct tatggtgcgg
     5641 atcgggatct tcagcgtgct ctacacggtt ccggccacca tcgtgatcgc ctgttacttc
     5701 tacgagcagg ccttccgcgc gcactgggag cagacgtgga gcgcgcactc gtgcaagaca
     5761 tacgcggtgc cgtgtcccgc gcacaacccg cagatgaacc cggacttcac ggtgttcatg
     5821 atcaagtacc tgatgaccct gatcgtgggc atcacctccg gcttctggat atggtctgga
     5881 aagacgctca attcctggag gaagttttac acgagactgg cgaacagcaa acagggagaa
     5941 accacagtgg gaggaggatc aggaggagga tcaatggtga gcaagggcga ggagaataac
     6001 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     6061 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgaggcctt tcagaccgct
     6121 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     6181 ttcatgtacg gctccaaggt ctacattaag cacccagccg acatccccga ctacttcaag
     6241 ctgtccttcc ccgagggctt caggtgggag cgcgtgatga acttcgagga cggcggcatt
     6301 attcacgtta accaggactc ctccctgcag gacggcgtgt tcatctacaa ggtgaagctg
     6361 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     6421 gcctccgagg agcggatgta ccccgaggac ggcgccctga agagcgagat caagaagagg
     6481 ctgaagctga aggacggcgg ccactacgcc gccgaggtca agaccaccta caaggccaag
     6541 aagcccgtgc agctgcccgg cgcctacatc gtcgacatca agttggacat cgtgtcccac
     6601 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     6661 ggcatggacg agctgtacaa gtaagcggcc gcaaacaaga atctctagtt ttctttcttg
     6721 cttttacttt tacttcctta atactcaagt acaattttaa tggagtactt ttttactttt
     6781 actcaagtaa gattctagcc agatactttt acttttaatt gagtaaaatt ttccctaagt
     6841 acttgtactt tcacttgagt aaaatttttg agtacttttt acacctctgc tcgaccatat
     6901 gggagagctc ccaacgcgtt ggatgcatag cttgagtatt ctatagtgtc acctaaatag
     6961 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7021 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     7081 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7141 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     7201 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7261 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7321 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7381 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7441 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7501 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7561 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7621 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7681 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7741 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7801 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7861 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7921 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7981 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     8041 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     8101 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     8161 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     8221 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     8281 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     8341 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8401 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     8461 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     8521 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     8581 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8641 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8701 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     8761 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8821 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8881 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8941 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     9001 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     9061 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     9121 gccacctgat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga
     9181 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     9241 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     9301 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     9361 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     9421 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     9481 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     9541 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     9601 acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac
     9661 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     9721 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     9781 acgacggcca gtgaattgta atacgactca ctata
//
