LOCUS       sequence-48260-e        4464 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 4464)
  AUTHORS   Joyce PI, Satija R, Chen M, Kuwabara PE
  TITLE     The atypical calpains: evolutionary analyses and roles in 
            Caenorhabditis elegans cellular degeneration.
  JOURNAL   PLoS Genet. 2012 Mar;8(3):e1002602. Epub 2012 Mar 29.
  PUBMED    22479198
REFERENCE   2  (bases 1 to 4464)
  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: "PLoS 
            Genet."; date: "2012-03"; volume: "8(3)"; pages: "e1002602. Epub 
            2012 Mar 29"
COMMENT     Sequence Label: pPJ-1
FEATURES             Location/Qualifiers
     source          1..4464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             146..166
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             188..208
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             230..250
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             272..292
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             305..982
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             305..982
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(435..453)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     703..722
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     897..920
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1904..1926)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2026..2045
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2123..2141)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2209..2313
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2314..2382,2383..3174)
                     /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(2532..2551)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3345..3933
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3834..3853
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4087..4104
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4221..4242
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4257..4262,4263..4280,4281..4287)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4295..4311
                     /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     4300..4322
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4319..4335
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4319..4335
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatccccgg gattggccaa aggacccaaa
       61 ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc cttggagggt
      121 accgagctca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg cagaaaaaat
      181 gactgctcca aagaagaagc gtaaggtacc ggcagaaaaa atgactgctc caaagaagaa
      241 gcgtaaggta ccggcagaaa aaatgactgc tccaaagaag aagcgtaagg taccggtaga
      301 aaaaatggcc tcctccgagg acgtcatcaa ggagttcatg cgcttcaagg tgcgcatgga
      361 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
      421 gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga
      481 catcctgtcc cctcagttcc agtacggctc caaggcctac gtgaagcacc ccgccgacat
      541 ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
      601 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat
      661 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa
      721 gaccatgggc tgggaggcct ccaccgagcg gatgtacccc gaggacggcg ccctgaaggg
      781 cgagatcaag atgaggctga agctgaagga cggcggccac tacgacgccg aggtcaagac
      841 cacctacatg gccaagaagc ccgtgcagct gcccggcgcc tacaagaccg acatcaagct
      901 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgagc gcgccgaggg
      961 ccgccactcc accggcgcct aagctagcga attccaactg agcgccggtc gctaccatta
     1021 ccaacttgtc tggtgtcaaa aataataggg gccgctgtca tcagagtaag tttaaactga
     1081 gttctactaa ctaacgagta atatttaaat tttcagcatc tcgcgcccgt gcctctgact
     1141 tctaagtcca attactcttc aacatcccta catgctcttt ctccctgtgc tcccaccccc
     1201 tatttttgtt attatcaaaa aacttctctt aatttctttg ttttttagct tcttttaagt
     1261 cacctctaac aatgaaattg tgtagattca aaaatagaat taattcgtaa taaaaagtcg
     1321 aaaaaaattg tgctccctcc ccccattaat aataattcta tcccaaaatc tacacaatgt
     1381 tctgtgtaca cttcttatgt tttttacttc tgataaattt ttttgaaaca tcatagaaaa
     1441 aaccgcacac aaaatacctt atcatatgtt acgtttcagt ttatgaccgc aatttttatt
     1501 tcttcgcacg tctgggcctc tcatgacgtc aaatcatgct catcgtgaaa aagttttgga
     1561 gtatttttgg aatttttcaa tcaagtgaaa gtttatgaaa ttaattttcc tgcttttgct
     1621 ttttggggtt tcccctattg tttgtcaaga tttcgaggac ggcgtttttc ttgctaaaat
     1681 cacaagtatt gatgagcacg atgcaagaaa gatcggaaga aggtttgggt ttgaggctca
     1741 gtggaaggtg agtagaagtt gataatttga aagtggagta gtgtctatgg ggtttttgcc
     1801 ttaaatgaca gaatacattc ccaatatacc aaacataact gtttcctact agtcggccgt
     1861 acgggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     1921 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     1981 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     2041 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     2101 accgcatcag gcggccttaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     2161 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2221 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2281 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     2341 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     2401 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     2461 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     2521 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     2581 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     2641 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     2701 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     2761 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     2821 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     2881 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2941 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3001 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     3061 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3121 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     3181 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3241 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3301 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3361 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3421 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3481 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3541 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3601 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3661 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3721 tacctacagc gtgagcattg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3781 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3841 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3901 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3961 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4021 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4081 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     4141 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     4201 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     4261 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     4321 ggaaacagct atgaccatga ttacgccaag ctgtaagttt aaacatgatc ttactaacta
     4381 actattctca tttaaatttt cagagcttaa aaatggctga aatcactcac aacgatggat
     4441 acgctaacaa cttggaaatg aaat
//
