LOCUS       sequence-148056-        3981 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 3981)
  AUTHORS   Clipstone NA, Crabtree GR
  TITLE     Identification of calcineurin as a key signalling enzyme in 
            T-lymphocyte activation.
  JOURNAL   Nature. 1992 Jun 25. 357(6380):695-7.
  PUBMED    1377362
REFERENCE   2  (bases 1 to 3981)
  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. 
            1992 Jun 25. 357(6380):695-7."
COMMENT     Sequence Label: pBJ5 mCnB
FEATURES             Location/Qualifiers
     source          1..3981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..330,331..345,346..614)
                     /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      181..316
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     LTR             346..614
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     intron          658..754
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     polyA_signal    1386..1520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1410..1429)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1464..1483
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1663..1685)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1771..1911
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1826..1845
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1926..1943)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2097..2685)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2177..2196)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2856..3647,3648..3716))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3479..3498
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3717..3821)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3889..3907
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(3977..3981,1..16))
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
       61 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
      121 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
      181 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
      241 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
      301 ggcttttttg gaggcctagg cttttgcaaa aagctcctcg atcgaggggc tcgcatctct
      361 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
      421 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
      481 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
      541 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg
      601 cgccgttaca gatccaagct cctcgatcga ggaactgaaa aaccagaaag ttaactggta
      661 agtttagtct ttttgtcttt tatttcaggt cccggatccg gtggtggtgc aaatcaaaga
      721 actgctcctc agtggatgtt gcctttactt ctaggcctgt acggaagtgt tacttctgct
      781 ctaaaagctg ctctagccct cgagggctag agcggccgcg gaggccgaat tgatccccat
      841 gggaaatgag gcaagttacc ctttggaaat gtgctcacac tttgatgctg atgaaattaa
      901 aaggctagga aagagattca agaagcttga tttggacaat tctggttctt tgagcgtgga
      961 agagttcatg tctctgcctg agttacagca gaacccttta gtacagcggg taatagatat
     1021 attcgacaca gacggcaacg gagaagtgga cttcaaagaa ttcattgaag gagtgtctca
     1081 gttcagtgtc aaaggcgata aggaacagaa gttgaggttt gcttttcgta tctatgacat
     1141 ggataaagat ggctatattt ccaatggaga actcttccag gtgttgaaga tgatggtggg
     1201 caacaatctg aaagatacac aattacagca gattgtagac aaaaccataa taaatgcaga
     1261 taaggatgga gatggaagaa tatcctttga ggaattctgt gctgtcgtag gtggcctaga
     1321 tatccacaaa aagatggtgg tagatgtgtg actcttatcg ggtacaattc ctgattgatt
     1381 gaccggatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     1441 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     1501 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     1561 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt
     1621 atgatccggc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     1681 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     1741 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     1801 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     1861 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc
     1921 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     1981 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2041 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2101 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2161 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2221 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2281 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2341 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2401 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2461 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2521 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2581 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2641 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2701 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     2761 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     2821 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     2881 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     2941 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3001 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3061 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3121 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc
     3181 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3241 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3301 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3361 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3421 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat
     3481 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     3541 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     3601 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     3661 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     3721 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     3781 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     3841 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     3901 acgaggccct ttcgtcttca agaattggtc gaccaattct catgtttgac agcttatcat
     3961 cgataagctt ggctgtggaa t
//
