LOCUS       sequence-469389-        6738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6738)
  AUTHORS   Hedgepeth CM, Conrad LJ, Zhang J, Huang HC, Lee VM, Klein PS
  TITLE     Activation of the Wnt signaling pathway: a molecular mechanism for 
            lithium action.
  JOURNAL   Dev Biol. 1997 May 1. 185(1):82-91.
  PUBMED    9169052
REFERENCE   2  (bases 1 to 6738)
  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: "Dev Biol. 
            1997 May 1. 185(1):82-91."
COMMENT     Sequence Label: tau/pET29b
FEATURES             Location/Qualifiers
     source          1..6738
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     563..582
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(602..679)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     885..904
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     992..1011
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        992..1010
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1011..1035
                     /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)."
     RBS             1050..1072
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2590..2607
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2660..2678)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2674..2721
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2758..3213
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2845..2864)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3055..3076
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3305..4120)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3305..4120)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4028..4047
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4112..4131
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4242..4830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4731..4750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4984..5001
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5016..5155
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5081..5100)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5241..5263
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5257..5448)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6257..6738,1..601))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
       61 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
      121 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
      181 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
      241 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
      301 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
      361 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
      421 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
      481 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
      541 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
      601 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
      661 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
      721 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
      781 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
      841 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
      901 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
      961 aggatcgaga tcgatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga
     1021 gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga gatatacata
     1081 tggctgagcc ccgccaggag ttcgaagtga tggaagatca cgctgggacg tacgggttgg
     1141 gggacaggaa agatcagggg ggctacacca tgcaccaaga ccaagagggt gacacggacg
     1201 ctggcctgaa agaatctccc ctgcagaccc ccactgagga cggatctgag gaaccgggct
     1261 ctgaaacctc tgatgctaag agcactccaa cagcggaaga tgtgacagca cccttagtgg
     1321 atgagggagc tcccggcaag caggctgccg cgcagcccca cacggagatc ccagaaggaa
     1381 ccacagctga agaagcaggc attggagaca cccccagcct ggaagacgaa gctgctggtc
     1441 acgtgaccca agctcgcatg gtcagtaaaa gcaaagacgg gactggaagc gatgacaaaa
     1501 aagccaaggg ggctgatggt aaaacgaaga tcgccacacc gcggggagca gcccctccag
     1561 gccagaaggg ccaggccaac gccaccagga ttccagcaaa aaccccgccc gctccaaaga
     1621 caccacccag ctctggtgaa cctccaaaat caggggatcg cagcggctac agcagccccg
     1681 gctccccagg cactcccggc agccgctccc gcaccccgtc ccttccaacc ccacccaccc
     1741 gggagcccaa gaaggtggca gtggtccgta ctccacccaa gtcgccgtct tccgccaaga
     1801 gccgcctgca gacagccccc gtgcccatgc cagacctgaa gaatgtcaag tccaagatcg
     1861 gctccactga gaacctgaag caccagccgg gaggcgggaa ggtgcagata attaataaga
     1921 agctggatct tagcaacgtc cagtccaagt gtggctcaaa ggataatatc aaacacgtcc
     1981 cgggaggcgg cagtgtgcaa atagtctaca aaccagttga cctgagcaag gtgacctcca
     2041 agtgtggctc attaggcaac atccatcata aaccaggagg tggccaggtg gaagtaaaat
     2101 ctgagaagct tgacttcaag gacagagtcc agtcgaagat tgggtccctg gacaatatca
     2161 cccacgtccc tggcggagga aataaaaaga ttgaaaccca caagctgacc ttccgcgaga
     2221 acgccaaagc caagacagac cacggggcgg agatcgtgta caagtcgcca gtggtgtctg
     2281 gggacacgtc tccacggcat ctcagcaatg tctcctccac cggcagcatc gacatggtag
     2341 actcgcccca gctcgccacg ctagctgacg aggtgtctgc ctccctggcc aagcagggtt
     2401 tgtgatcagg cccctggggc ggtcaataat tgtggagagg agagaatgag agagtgtgga
     2461 aaaaaaaaga ataatgaccc ggcccccgcc ctctgccccc agctgctcct cgcagttcgg
     2521 gaattagcta agaaggagat ataggatccg aattcgagct ccgtcgacaa gcttgcggcc
     2581 gcactcgagc accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa
     2641 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     2701 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg
     2761 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2821 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2881 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2941 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3001 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3061 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3121 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3181 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     3241 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta
     3301 attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     3361 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     3421 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     3481 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     3541 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
     3601 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     3661 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     3721 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     3781 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     3841 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     3901 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     3961 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     4021 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     4081 gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     4141 ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc
     4201 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4261 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4321 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4381 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4441 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4501 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4561 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4621 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4681 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4741 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4801 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4861 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4921 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4981 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     5041 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg
     5101 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     5161 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     5221 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     5281 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     5341 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     5401 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     5461 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     5521 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     5581 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     5641 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
     5701 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
     5761 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     5821 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     5881 cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc
     5941 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
     6001 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
     6061 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
     6121 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     6181 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     6241 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6301 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     6361 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     6421 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     6481 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatgt
     6541 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     6601 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     6661 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     6721 gagtgagata tttatgcc
//
