LOCUS       sequence-47286-e        4720 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 4720)
  AUTHORS   Graef IA, Mermelstein PG, Stankunas K, Neilson JR, Deisseroth K, 
            Tsien RW, Crabtree GR
  TITLE     L-type calcium channels and GSK-3 regulate the activity of NF-ATc4 
            in hippocampal neurons.
  JOURNAL   Nature. 1999 Oct 14. 401(6754):703-8.
  PUBMED    10537109
REFERENCE   2  (bases 1 to 4720)
  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: "Nature. 
            1999 Oct 14. 401(6754):703-8."
COMMENT     Sequence Label: pShGSK3bE
FEATURES             Location/Qualifiers
     source          1..4720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..31)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(16..345,346..361,362..631)
                     /label=SR-alpha promoter
                     /note="hybrid promoter consisting of the SV40 early 
                     promoter plus part of the long terminal repeat of human 
                     T-lymphotrophic virus 1"
     rep_origin      196..331
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     LTR             362..631
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     intron          676..769
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             1649..1660
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             2098..2124
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2155..2289
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2179..2198)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2233..2252
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2429..2451)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2537..2677
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2592..2611
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2692..2709)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2863..3451)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2943..2962)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3622..4413,4414..4482))
                     /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     4245..4264
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4483..4587)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4655..4673
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aagcttctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
       61 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
      121 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
      181 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
      241 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
      301 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cctccgatcg
      361 aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccaacgcc
      421 ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct
      481 aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct tggagcctac
      541 ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc tacgtctttg
      601 tttcgttttc tgttctgcgc cgttacagat ccaagctgcc tcgatcgagg aactgaaaaa
      661 ccagaaagtt aactggtaag tttagtcttt ttgtctttta tttcaggtcc cggatccggt
      721 ggtgcaaatc aaagaactgc tcctcagtgg atgttgcctt tacttctagg cctgtacgga
      781 ggtgttactt ctgctctaaa agctgctcta gccgcggccg cgatgctcga gatgtcaggg
      841 cggcccagaa ccacctcctt tgcggagagc tgcaagccgg tgcagcagcc ttcagctttt
      901 ggcagcatga aagttagcag agacaaggac ggcagcaagg tgacaacagt ggtggcaact
      961 cctgggcagg gtccagacag gccacaagaa gtcagctata cagacactaa agtgattgga
     1021 aatggatcat ttggtgtggt atatcaagcc aaactttgtg attcaggaga actggtcgcc
     1081 atcaagaaag tattgcagga caagagattt aagaatcgag agctccagat catgagaaag
     1141 ctagatcact gtaacatagt ccgattgcgt tatttcttct actccagtgg tgagaagaaa
     1201 gatgaggtct atcttaatct ggtgctggac tatgttccgg aaacagtata cagagttgcc
     1261 agacactata gtcgagccaa acagacgctc cctgtgattt atgtcaagtt gtatatgtat
     1321 cagctgttcc gaagtttagc ctatatccat tcctttggaa tctgccatcg ggatattaaa
     1381 ccgcagaacc tcttgttgga tcctgatact gctgtattaa aactctgtga ctttggaagt
     1441 gcaaagcagc tggtccgagg agaacccaat gtttcgtata tctgttctcg gtactatagg
     1501 gcaccagagt tgatctttgg agccactgat tatacctcta gtatagatgt atggtctgct
     1561 ggctgtgtgt tggctgagct gttactagga caaccaatat ttccagggga tagtggtgtg
     1621 gatcagttgg tagaaataat caaggtcctg ggaactccaa caagggagca aatcagagaa
     1681 atgaacccaa actacacaga atttaaattc cctcaaatta aggcacatcc ttggactaag
     1741 gtcttccgac cccgaactcc accggaggca attgcactgt gtagccgtct gctggagtat
     1801 acaccaactg cccgactaac accactggaa gcttgtgcac attcattttt tgatgaatta
     1861 cgggacccaa atgtcaaact accaaatggg cgagacacac ctgcactctt caacttcacc
     1921 actcaagaac tgtcaagtaa tccacctctg gctaccatcc ttattcctcc tcatgctcgg
     1981 attcaagcag ctgcttcaac ccccacaaat gccacagcag cgtcagatgc taatactgga
     2041 gaccgtggac agaccaataa tgctgcttct gcatcagctt ccaactccac cgtcgactat
     2101 ccgtacgacg taccagacta cgcactcgac taagaattcc tgattgattg accggatcca
     2161 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     2221 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     2281 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
     2341 gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta tgatcctgcc
     2401 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     2461 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     2521 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
     2581 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     2641 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
     2701 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2761 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2821 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2881 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2941 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3001 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     3061 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3121 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3181 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3241 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3301 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3361 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3421 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3481 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3541 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3601 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3661 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3721 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3781 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3841 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3901 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3961 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4021 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4081 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4141 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4201 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4261 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4321 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4381 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4441 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4501 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4561 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     4621 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     4681 tcttcaagat cgacctcatg tttgacagct tatcatcgat
//
