LOCUS       sequence-142354-        8422 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 8422)
  AUTHORS   Lancaster MA, Louie CM, Silhavy JL, Sintasath L, Decambre M, Nigam 
            SK, Willert K, Gleeson JG
  TITLE     Impaired Wnt-beta-catenin signaling disrupts adult renal homeostasis
            and leads to cystic kidney ciliopathy.
  JOURNAL   Nat Med. 2009 Sep . 15(9):1046-54.
  PUBMED    19718039
REFERENCE   2  (bases 1 to 8422)
  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: "Nat Med. 
            2009 Sep . 15(9):1046-54."
COMMENT     Sequence Label: EGFP-Ahi1
FEATURES             Location/Qualifiers
     source          1..8422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      614..623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(620..622,623..625,626..1336)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(620..622,623..625,626..1336)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(665..686)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(926..945)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1273..1294
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5217..5338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5254..5273)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5308..5327
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5345..5800)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5482..5503)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5694..5713
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5827..5931
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5933..6290
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5933..6124
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5956..5976)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6141..6276
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6325..7119
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6325..7119
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6379..6398)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6989..7008
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7307..7326)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7351..7398
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7727..8315
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8216..8235
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
      661 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
      721 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
      781 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
      841 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
      901 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
      961 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1021 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     1081 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     1141 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1201 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     1261 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1321 ggacgagctg tacaagtact cagatctcga gctcggatcc actagtaacg gccgccagtg
     1381 tgctggaatt cgccctttgc ctacagctga gagtgaagca aaagtaaaaa ccaaagttcg
     1441 ctttgaagaa ttgcttaaga cccacagtga tctaatgcgt gaaaagaaaa aactgaagaa
     1501 aaaacttgtc aggtctgaag aaaacatctc acctgacact attagaagca atcttcacta
     1561 tatgaaagaa actacaagtg atgatcccga cactattaga agcaatcttc cccatattaa
     1621 agaaactaca agtgatgatg taagtgctgc taacactaac aacctgaaga agagcacgag
     1681 agtcactaaa aacaaattga ggaacacaca gttagcaact gaaaatccta atggtgatgc
     1741 tagtgtagag gaagacaaac aaggaaagcc aaataaaaag gtgataaaga cggtgcccca
     1801 gttgactaca caagacctga aaccggaaac tcctgagaat aaggttgatt ctacacacca
     1861 gaaaacacat acaaagccac agccaggcgt tgatcatcag aaaagtgaga aggcaaatga
     1921 gggaagagaa gagactgatt tagaagagga tgaagaattg atgcaagcat atcagtgcca
     1981 tgtaactgaa gaaatggcaa aggagattaa gaggaaaata agaaagaaac tgaaagaaca
     2041 gttgacttac tttccctcag atactttatt ccatgatgac aaactaagca gtgaaaaaag
     2101 gaaaaagaaa aaggaagttc cagtcttctc taaagctgaa acaagtacat tgaccatctc
     2161 tggtgacaca gttgaaggtg aacaaaagaa agaatcttca gttagatcag tttcttcaga
     2221 ttctcatcaa gatgatgaaa taagctcaat ggaacaaagc acagaagaca gcatgcaaga
     2281 tgatacaaaa cctaaaccaa aaaaaacaaa aaagaagact aaagcagttg cagataataa
     2341 tgaagatgtt gatggtgatg gtgttcatga aataacaagc cgagatagcc cggtttatcc
     2401 caaatgtttg cttgatgatg accttgtctt gggagtttac attcaccgaa ctgatagact
     2461 taagtcagat tttatgattt ctcacccaat ggtaaaaatt catgtggttg atgagcatac
     2521 tggtcaatat gtcaagaaag atgatagtgg acggcctgtt tcatcttact atgaaaaaga
     2581 gaatgtggat tatattcttc ctattatgac ccagccatat gattttaaac agttaaaatc
     2641 aagacttcca gagtgggaag aacaaattgt atttaatgaa aattttccct atttgcttcg
     2701 aggctctgat gagagtccta aagtcatcct gttctttgag attcttgatt tcttaagcgt
     2761 ggatgaaatt aagaataatt ctgaggttca aaaccaagaa tgtggctttc ggaaaattgc
     2821 ctgggcattt cttaagcttc tgggagccaa tggaaatgca aacatcaact caaaacttcg
     2881 cttgcagcta tattacccac ctactaagcc tcgatcccca ttaagtgttg ttgaggcatt
     2941 tgaatggtgg tcaaaatgtc caagaaatca ttacccatca acactgtacg taactgtaag
     3001 aggactgaaa gttccagact gtataaagcc atcttaccgc tctatgatgg ctcttcagga
     3061 ggaaaaaggt aaaccagtgc attgtgaacg tcaccatgag tcaagctcag tagacacaga
     3121 acctggatta gaagagtcaa aggaagtaat aaagtggaaa cgactccctg ggcaggcttg
     3181 ccgtatccca aacaaacacc tcttctcact aaatgcagga gaacgaggat gtttttgtct
     3241 tgatttctcc cacaatggaa gaatattagc agcagcttgt gccagccggg atggatatcc
     3301 aattatttta tatgaaattc cttctggacg tttcatgaga gaattgtgtg gccacctcaa
     3361 tatcatttat gatctttcct ggtcaaaaga tgatcactac atccttactt catcatctga
     3421 tggcactgcc aggatatgga aaaatgaaat aaacaataca aatactttca gagttttacc
     3481 tcatccttct tttgtttaca cggctaaatt ccatccagct gtaagagagc tagtagttac
     3541 aggatgctat gattccatga tacggatatg gaaagttgag atgagagaag attctgccat
     3601 attggtccga cagtttgatg ttcacaaaag ttttatcaac tcactttgtt ttgatactga
     3661 aggtcatcat atgtattcag gagattgtac aggggtgatt gttgtttgga atacctatgt
     3721 caagattaat gatttggaac attcagtgca ccactggact ataaataagg aaattaaaga
     3781 aactgagttt aagggaattc caataagtta tttggagatt catcccaatg gaaaacgttt
     3841 gttaatccat accaaagaca gtactttgag aattatggat ctccggatat tagtagcaag
     3901 gaagtttgta ggagcagcaa attatcggga gaagattcat agtactttga ctccatgtgg
     3961 gacttttctg tttgctggaa gtgaggatgg tatagtgtat gtttggaacc cagaaacagg
     4021 agaacaagta gccatgtatt ctgacttgcc attcaagtca cccattcgag acatttctta
     4081 tcatccattt gaaaatatgg ttgcattctg tgcatttggg caaaatgagc caattcttct
     4141 gtatatttac gatttccatg ttgcccagca ggaggctgaa atgttcaaac gctacaatgg
     4201 aacatttcca ttacctggaa tacaccaaag tcaagatgcc ctatgtacct gtccaaaact
     4261 accccatcaa ggctcttttc agattgatga atttgtccac actgaaagtt cttcaacgaa
     4321 gatgcagcta gtaaaacaga ggcttgaaac tgtcacagag gtgatacgtt cctgtgctgc
     4381 aaaagtcaac aaaaatctct catttacttc accaccagca gtttcctcac aacagtctaa
     4441 gttaaagcag tcaaacatgc tgaccgctca agagattcta catcagtttg gtttcactca
     4501 gaccgggatt atcagcatag aaagaaagcc ttgtaaccat caggtagata cagcaccaac
     4561 ggtagtggct ctttatgact acacagcgaa tcgatcagat gaactaacca tccatcgcgg
     4621 agacattatc cgagtgtttt tcaaagataa tgaagactgg tggtatggca gcataggaaa
     4681 gggacaggaa ggttattttc cagctaatca tgtggctagt gaaacactgt atcaagaact
     4741 gcctcctgag ataaaggagc gatcccctcc tttaagccct gaggaaaaaa ctaaaataga
     4801 aaaatctcca gctcctcaaa agcaatcaat caataagaac aagtcccagg acttcagact
     4861 aggctcagaa tctatgacac attctgaaat gagaaaagaa cagagccatg aggaccaagg
     4921 acacataatg gatacacgga tgaggaagaa caagcaagca ggcagaaaag tcactctaat
     4981 agagtaaaga attgaagaaa agttaagagc tgccgaaatg cacaagggcg aattctgcag
     5041 atatccatca cactggcggc cgctcgagca tgcatctaga gggcccggga tccaccggat
     5101 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     5161 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     5221 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5281 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac
     5341 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     5401 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     5461 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     5521 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     5581 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     5641 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     5701 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     5761 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     5821 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5881 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     5941 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     6001 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     6061 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     6121 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     6181 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     6241 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     6301 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     6361 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     6421 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     6481 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     6541 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     6601 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     6661 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     6721 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     6781 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     6841 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     6901 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     6961 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     7021 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     7081 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     7141 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     7201 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     7261 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     7321 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     7381 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     7441 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     7501 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     7561 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     7621 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     7681 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     7741 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     7801 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     7861 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     7921 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     7981 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     8041 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     8101 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     8161 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     8221 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     8281 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     8341 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     8401 ctgtggataa ccgtattacc gc
//
