LOCUS       sequence-11461-e        5682 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 5682)
  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 5682)
  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: pBS_hNFATc4nuc
FEATURES             Location/Qualifiers
     source          1..5682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2713..2730
                     /codon_start=1
                     /product="epitope tag from middle T antigen of mouse 
                     polyoma virus"
                     /label=Glu-Glu tag
                     /translation="EYMPME"
     primer_bind     complement(2765..2784)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2766..2784)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2794..2811)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2794..2810)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2803..2825)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2952..3407
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3039..3058)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3249..3270
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3433..3537
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3538..3606,3607..4398)
                     /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     complement(3756..3775)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4569..5157
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5058..5077
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5311..5328
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5445..5466
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5481..5486,5487..5504,5505..5511)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5519..5535
                     /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)."
     primer_bind     5524..5546
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5543..5559
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5543..5559
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5578..5598
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5580..5598
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     5627..5643
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5628..5644
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 atgggggcgg ccagctgcga ggatgaggag ctggaattta agctggtgtt cggggaggaa
       61 aaggaggccc ccccgctggg cgcgggggga ttgggggaag aactggactc agaggatgcc
      121 ccgccatgct gccgtctggc cttgggagag ccccctccct atggcgctgc acctatcggt
      181 attccccgac ctccaccccc tcggcctggc atgcattcgc caccgccgcg accagccccc
      241 tcacctggca cctgggagag ccagcccgcc aggtcggtga ggctgggagg accaggaggg
      301 ggtgctgggg gtgctggggg tggccgtgtt ctcgagtgtc ccagcatccg catcacctcc
      361 atctctccca cgccggagcc gccagcagcg ctggaggaca accctgatgc ctggggggac
      421 ggctctccta gagattaccc cccaccagaa ggctttgggg gctacagaga agcaggggcc
      481 cagggtgggg gggccttctt cgccccagcc cctggcgccg ccgccctgtc ctcgtggagc
      541 ttcttctccg atgcctctga cgaggcagcc ctgtatgcag cctgcgacga ggtggagtct
      601 gagctaaatg aggcggcctc ccgctttggc ctgggcgccc cgctgcccgc cccccgggcc
      661 gcccctcggc catggacccc cgaagatccc tggagcctgt atggtccaag ccccggaggc
      721 cgagggccag aggatagctg gctactcctc agtgctcctg ggcccacccc agcctccccg
      781 cggcctgcct ctccatgtgg caagcggcgc tattccagct cgggaacccc atcttcagcc
      841 tccccagctc tgtcccgccg tggcagcctg ggggaagagg ggtctgagcc acctccacca
      901 cccccattgc ctctggcccg ggacccgggc tcccctggtc cctttgacta tgtgggggcc
      961 ccaccagctg agagcatccc tcagaagaca cggcggactt ccagcgagca ggcagtggct
     1021 ctgcctcggt ctgaggagcc tgcctcatgc aatgggaagc tgcccttggg agcagaggag
     1081 tctgtggctc ctccaggagg ttcccggaag gaggtggctg gcatggacta cctggcagtg
     1141 ccctccccac tcgcttggtc caaggcccgg attgggggac acagccctat cttcaggacc
     1201 tctgccctac ccccactgga ctggcctctg cccagccaat atgagcagct ggagctgagg
     1261 atcgaggtac agcctagagc ccaccaccgg gcccactatg agacagaagg cagccgtgga
     1321 gctgtcaaag ctgcccctgg cggtcacccc gtagtcaagc tcctaggcta cagtgagaag
     1381 ccactgaccc tacagatgtt catcggcact gcagatgaaa ggaacctgcg gcctcatgcc
     1441 ttctatcagg tgcaccgtat cacaggcaag atggtggcca cggccagcta tgaagccgta
     1501 gtcagtggca ccaaggtgtt ggagatgact ctgctgcctg agaacaacat ggcggccaac
     1561 attgactgcg cgggaatcct gaagcttcgg aattcagaca ttgagcttcg gaagggtgag
     1621 acggacatcg ggcgcaaaaa cacacgtgta cggctggtgt tccgggtaca cgtgccccag
     1681 ggcggcggga aggtcgtctc agtacaggca gcatcggtgc ccatcgagtg ctcccagcgc
     1741 tcagcccagg agctgcccca ggtggaggcc tacagcccca gtgcctgctc tgtgagagga
     1801 ggcgaggaac tggtactgac cggctccaac ttcctgccag actccaaggt ggtgttcatt
     1861 gagaggggtc ctgatgggaa gctgcaatgg gaggaggagg ccacagtgaa ccgactgcag
     1921 agcaacgagg tgacgctgac cctgactgtc cccgagtaca gcaacaagag ggtttcccgg
     1981 ccagtccagg tctactttta tgtctccaat gggcggagga aacgcagtcc tacccagagt
     2041 ttcaggtttc tgcctgtgat ctgcaaagag gagcccctac cggactcatc tctgcggggt
     2101 ttcccttcag catcggcaac cccctttggc actgacatgg acttctcacc acccaggccc
     2161 ccctacccct cctatcccca tgaagaccct gcttgcgaaa ctccttacct atcagaaggc
     2221 ttcggctatg gcatgccccc tctgtacccc cagacggggc ccccaccatc ctacagaccg
     2281 ggcctgcgga tgttccctga gactaggggt accacaggtt gtgcccaacc acctgcagtt
     2341 tccttccttc cccgcccctt ccctagtgac ccgtatggag ggcggggctc ctctttcccc
     2401 ctggggctgc cattctctcc gccagccccc tttcggccgc ctcctcttcc tgcatcccca
     2461 ccgcttgaag gccccttccc ttcccagagt gatgtgcatc ccctacctgc tgagggatac
     2521 aataaggtag ggccaggcta tggccctggg gagggggctc cggagcagga gaaatccagg
     2581 ggtggctaca gcagcggctt tcgagacagt gtccctatcc agggtatcac gctggaggaa
     2641 gtgagtgaga tcattggccg agacctgagt ggcttccctg cacctcctgg agaagagcct
     2701 cctgccggca gcgaatatat gccaatggaa tgagcggccg ccaccgcggt ggagctccaa
     2761 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     2821 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     2881 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     2941 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3001 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3061 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3121 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3181 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3241 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3301 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3361 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     3421 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3481 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3541 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3601 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3661 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3721 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3781 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3841 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3901 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3961 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4021 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4081 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4141 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4201 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4261 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4321 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4381 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4441 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4501 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4561 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4621 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4681 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     4741 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4801 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4861 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4921 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4981 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5041 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5101 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5161 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5221 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     5281 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     5341 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     5401 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     5461 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     5521 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
     5581 attaaccctc actaaaggga acaaaagctg ggtaccgggc cccccctcga ggtcgacggt
     5641 atcgataagc ttgatatcga attcctgcag cccgggggat cc
//
