LOCUS       sequence-257353-        7697 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7697)
  AUTHORS   Wrobel CN, Mutch CA, Swaminathan S, Taketo MM, Chenn A
  TITLE     Persistent expression of stabilized beta-catenin delays maturation 
            of radial glial cells into intermediate progenitors.
  JOURNAL   Dev Biol. 2007 Sep 15. 309(2):285-97.
  PUBMED    17706960
REFERENCE   2  (bases 1 to 7697)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Dev Biol. 
            2007 Sep 15. 309(2):285-97."
COMMENT     Sequence Label: pCAG-delta90GFP
FEATURES             Location/Qualifiers
     source          1..7697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          165..1181
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1105..1127
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1189..1208
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1314..1323
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3408..3417
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3414..3416,3417..3419,3420..4133)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3459..3480)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3720..3739)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4067..4088
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4220..4239)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4285..4340
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4339..4358)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4701..4717)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4701..4717)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4714..4736)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4725..4741
                     /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(4749..4755,4756..4773,4774..4779))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4794..4815
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4874..5069
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4920..5055
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5075..5209
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5112..5131)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5166..5185
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5277..5294)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5448..6036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5528..5547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6207..6998,6999..7067))
                     /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     6830..6849
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7068..7172)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        7203..7582
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7584..7697,1..164)
                     /label=chicken beta-actin promoter
ORIGIN
        1 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
       61 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
      121 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct
      181 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
      241 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
      301 gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc
      361 gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg
      421 ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg
      481 ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt
      541 gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag
      601 cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct ccccgagttg
      661 ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg
      721 tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg
      781 gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg
      841 cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt
      901 cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct agcgggcgcg
      961 gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc
     1021 cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg gacggctgcc
     1081 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag
     1141 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
     1201 ttattgtgct gtctcatcat tttggcaaag aattctgcag tcgacggtac cgagctcgga
     1261 tccactagta acggccgcca gtgtgctgga attcccatcg attctagacc gcggccacca
     1321 tggctcagag ggtccgagct gccatgttcc ctgagacgct agatgagggc atgcagatcc
     1381 catccacgca gtttgacgct gctcatccca ctaatgtcca gcgcttggct gaaccatcac
     1441 agatgttgaa acatgcagtt gtcaatttga ttaactatca ggatgacgcg gaacttgcca
     1501 cacgtgcaat tcctgagctg acaaaactgc taaacgatga ggaccaggtg gtagttaata
     1561 aagctgctgt tatggtccat cagctttcca aaaaggaagc ttccagacat gccatcatgc
     1621 gctcccctca gatggtgtct gccattgtac gcaccatgca gaatacaaat gatgtagaga
     1681 cagctcgttg tactgctggg actctgcaca acctttctca ccaccgcgag ggcttgctgg
     1741 ccatctttaa gtctggtggc atcccagcgc tggtgaaaat gcttgggtca ccagtggatt
     1801 ctgtactgtt ctacgccatc acgacactgc ataatctcct gctccatcag gaaggagcta
     1861 aaatggcagt gcgcctagct ggtggactgc agaaaatggt tgctttgctc aacaaaacaa
     1921 acgtgaaatt cttggctatt acaacagact gccttcagat cttagcttat ggcaatcaag
     1981 agagcaagct catcattctg gccagtggtg gaccccaagc cttagtaaac ataatgagga
     2041 cctacactta tgagaagctt ctgtggacca caagcagagt gctgaaagtg ctgtctgtct
     2101 gctctagcaa caagccggcc attgtagaag ctggtgggat gcaggcactg gggcttcatc
     2161 tgacagaccc aagtcagcga cttgttcaaa actgtctttg gactctcaga aacctttcag
     2221 atgcagcgac taagcaggaa gggatggaag gcctccttgg gactctagtg cagcttctgg
     2281 gttccgatga tataaatgtg gtcacctgtg cagctggaat tctctctaac ctcacttgca
     2341 ataattacaa aaacaagatg atggtgtgcc aagtgggtgg catagaggct cttgtacgca
     2401 ccgtccttcg tgctggtgac agggaagaca tcactgagcc tgccatctgt gctcttcgtc
     2461 atctgaccag ccggcatcag gaagccgaga tggcccagaa tgccgttcgc cttcattatg
     2521 gactgcctgt tgtggttaaa ctcctgcacc caccatccca ctggcctctg ataaaggcaa
     2581 ctgttggatt gattcgaaac cttgcccttt gcccagcaaa tcatgcgcct ttgcgggaac
     2641 agggtgctat tccacgacta gttcagctgc ttgtacgagc acatcaggac acccaacggc
     2701 gcacctccat gggtggaacg cagcagcagt ttgtggaggg cgtgcgcatg gaggagatag
     2761 tagaagggtg tactggagct ctccacatcc ttgctcggga cgttcacaac cggattgtaa
     2821 tccgaggact caataccatt ccattgtttg tgcagttgct ttattctccc attgaaaata
     2881 tccaaagagt agctgcaggg gtcctctgtg aacttgctca ggacaaggag gctgcagagg
     2941 ccattgaagc tgagggagcc acagctcccc tgacagagtt actccactcc aggaatgaag
     3001 gcgtggcaac atacgcagct gctgtcctat tccgaatgtc tgaggacaag ccacaggatt
     3061 acaagaagcg gctttcagtc gagctgacca gttccctctt caggacagag ccaatggctt
     3121 ggaatgagac tgcagatctt ggactggaca ttggtgccca gggagaagcc cttggatatc
     3181 gccaggatga tcccagctac cgttcttttc actctggtgg atacggccag gatgccttgg
     3241 ggatggaccc tatgatggag catgagatgg gtggccacca ccctggtgct gactatccag
     3301 ttgatgggct gcctgatctg ggacacgccc aggacctcat ggatgggctg cccccaggtg
     3361 atagcaatca gctggcctgg tttgatactg acctggatcc accggtcgcc accatggtga
     3421 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     3481 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     3541 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     3601 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     3661 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     3721 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     3781 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     3841 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     3901 aggcgaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact
     3961 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     4021 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     4081 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc
     4141 gcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg
     4201 ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc
     4261 cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt
     4321 ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc
     4381 agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca
     4441 aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat
     4501 tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt
     4561 tttctttaac atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct
     4621 cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc gacctgcagc
     4681 ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4741 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4801 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4861 tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4921 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4981 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     5041 gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg
     5101 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5161 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa
     5221 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5281 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5341 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5401 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5461 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5521 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5581 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5641 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5701 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5761 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5821 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5881 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5941 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6001 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6061 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6121 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6181 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6241 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6301 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6361 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6421 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6481 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6541 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6601 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6661 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6721 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6781 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6841 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6901 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6961 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7021 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7081 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7141 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctggg
     7201 tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     7261 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     7321 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     7381 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     7441 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     7501 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     7561 attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat
     7621 ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc
     7681 gatgggggcg ggggggg
//
