LOCUS       sequence-322384-        5570 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 5570)
  AUTHORS   Moyers JS, Bilan PJ, Zhu J, Kahn CR
  TITLE     Rad and Rad-related GTPases interact with calmodulin and 
            calmodulin-dependent protein kinase II.
  JOURNAL   J Biol Chem. 1997 May 2. 272(18):11832-9.
  PUBMED    9115241
REFERENCE   2  (bases 1 to 5570)
  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: "J Biol 
            Chem. 1997 May 2. 272(18):11832-9."
COMMENT     Sequence Label: pGEX Rad C249
FEATURES             Location/Qualifiers
     source          1..5570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(83..88,89..104,105..111)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    119..135
                     /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)."
     CDS             158..811
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     769..791
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             818..835
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1520..1542)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1687..1705)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1773..1877
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1878..1946,1947..2738)
                     /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     complement(2096..2115)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2909..3497
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3398..3417
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3651..3668
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3741..3818
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3819..4901
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(3838..3857)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4950..4955,4956..4973,4974..4980)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4988..5004
                     /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     5012..5028
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5012..5028
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5024..5551
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5044..5061)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5044..5060)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5053..5075)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cgctcaaggc gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg
       61 gcaaatattc tgaaatgagc tgttgacaat taatcatcgg ctcgtataat gtgtggaatt
      121 gtgagcggat aacaatttca cacaggaaac agtattcatg tcccctatac taggttattg
      181 gaaaattaag ggccttgtgc aacccactcg acttcttttg gaatatcttg aagaaaaata
      241 tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt
      301 gggtttggag tttcccaatc ttccttatta tattgatggt gatgttaaat taacacagtc
      361 tatggccatc atacgttata tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga
      421 gcgtgcagag atttcaatgc ttgaaggagc ggttttggat attagatacg gtgtttcgag
      481 aattgcatat agtaaagact ttgaaactct caaagttgat tttcttagca agctacctga
      541 aatgctgaaa atgttcgaag atcgtttatg tcataaaaca tatttaaatg gtgatcatgt
      601 aacccatcct gacttcatgt tgtatgacgc tcttgatgtt gttttataca tggacccaat
      661 gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag ctatcccaca
      721 aattgataag tacttgaaat ccagcaagta tatagcatgg cctttgcagg gctggcaagc
      781 cacgtttggt ggtggcgacc atcctccaaa atcggatctg gttccgcgtg gatccatgcc
      841 ggtggacgag cgcgacctgc aggcggcgct gaccccgggt gccctgacgg cggccgcggc
      901 cgggacgggg acccagggtc ccaggctgga ctggcccgag gactccgagg actcgctcag
      961 ctcagggggc agcgactcag acgagagcgt ttacaaggtg ctgctgctgg gggcgcccgg
     1021 cgtgggcaag agcgccctgg cgcgcatctt cggcggtgtg gaggacgggc ctgaagcaga
     1081 ggcagcaggg cacacctatg atcgctccat tgtagtggac ggagaagagg catcactcat
     1141 ggtctacgac atttgggagc aggacggggg ccgctggttg cccggccact gcatggccat
     1201 gggggatgcc tatgtcattg tgtactcagt gacggacaag ggcagcttcg agaaggcctc
     1261 agaactgcgg gtccagctgc ggcgtgcacg gcaaacagat gatgtgccca tcatcctcgt
     1321 gggcaacaag agcgacctgg tgcgctctcg tgaggtctcg gtggatgagg gccgggcctg
     1381 cgcggtggtc tttgactgca agttcattga gacatcagcg gcattgcacc acaatgtcca
     1441 ggcgctgttt gaaggtgtct agaattcatc gtgactgact gacgatctgc ctcgcgcgtt
     1501 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     1561 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     1621 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtataat tcttgaagac
     1681 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     1741 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1801 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1861 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     1921 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     1981 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2041 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2101 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     2161 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2221 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2281 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2341 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2401 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2461 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2521 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2581 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2641 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2701 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     2761 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     2821 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2881 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2941 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3001 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3061 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3121 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3181 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3241 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3301 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3361 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3421 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3481 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3541 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3601 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3661 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3721 tttcacaccg cataaattcc gacaccatcg aatggcgcaa aacctttcgc ggtatggcat
     3781 gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg
     3841 atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca
     3901 gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca
     3961 ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca
     4021 cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg
     4081 atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta
     4141 aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc
     4201 tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc
     4261 ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc
     4321 gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc
     4381 cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca
     4441 atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac
     4501 aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc
     4561 agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata
     4621 tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca
     4681 ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct
     4741 ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa
     4801 ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     4861 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta
     4921 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     4981 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgga
     5041 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     5101 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     5161 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc
     5221 accagaagcg gtgccggaaa gctggctgga gtgcgatctt cctgaggccg atactgtcgt
     5281 cgtcccctca aactggcaga tgcacggtta cgatgcgccc atctacacca acgtgaccta
     5341 tcccattacg gtcaatccgc cgtttgttcc cacggagaat ccgacgggtt gttactcgct
     5401 cacatttaat gttgatgaaa gctggctaca ggaaggccag acgcgaatta tttttgatgg
     5461 cgttggaatt agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
     5521 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt
//
