LOCUS       sequence-67926-e        5117 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 5117)
  AUTHORS   Rhyner JA, Koller M, Durussel-Gerber I, Cox JA, Strehler EE
  TITLE     Characterization of the human calmodulin-like protein expressed in 
            Escherichia coli.
  JOURNAL   Biochemistry. 1992 Dec 29;31(51):12826-32.
  PUBMED    1334432
REFERENCE   2  (bases 1 to 5117)
  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: 
            "Biochemistry."; date: "1992-12-29"; volume: "31(51)"; pages: 
            "12826-32"
COMMENT     Sequence Label: pKK233-hCALML3
FEATURES             Location/Qualifiers
     source          1..5117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(204..209,210..226,227..233)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    241..257
                     /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     246..268
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(852..869)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1022..1108
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1200..1227
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1247..1338
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1339..1407,1408..2199)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1557..1576)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2370..2958
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2859..2878
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3112..3129
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3144..3284
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3210..3229)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3370..3392
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3386..3577)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5035..5054
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ggaattctca tgtttgacag cttatcatcg actgcacggt gcaccaatgc ttctggcgtc
       61 aggcagccat cggaagctgt ggattggctg tgcaggtcgt aaatcactgc ataattcgtg
      121 tcgctcaagg cgcactcccg ttctggataa tgttttttgc gccgacatat aaacggttct
      181 ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcc ggctcgtata atgtgtggaa
      241 ttgtgagcgg ataacaattt cacacaggaa acagaccatg gccgaccagc tgactgagga
      301 gcaggtcaca gaattcaagg aggccttctc cctgtttgac aaggatgggg acggctgcat
      361 caccacccgc gagctgggca cggtcatgcg gtccctgggc cagaacccca cggaggccga
      421 gctgcgggac atgatgagtg agatcgaccg ggacggcaac ggcaccgtgg acttccccga
      481 gttcctgggc atgatggcca ggaagatgaa ggacacggac aacgaggagg agatccgcga
      541 ggccttccgc gtgttcgaca aggacggcaa cggcttcgtc agcgccgccg agctgcgaca
      601 cgtcatgacc cggctggggg agaagctgag tgacgaggag gtggacgaga tgatccgggc
      661 cgcggacacg gacggagacg gacaggtgaa ctacgaggag tttgtccgtg tgctggtgtc
      721 caagtgaggc cggcgcccac catgctcctg ggcccccacg cggcccacag ggcaagaacc
      781 cggggatcct ctagagtcga cctgcaggca tgcaagcttg gctgttttgg cggatgagag
      841 aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat
      901 ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa
      961 cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
     1021 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     1081 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     1141 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     1201 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actctttttg tttatttttc
     1261 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1321 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1381 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1441 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1501 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1561 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     1621 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1681 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1741 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     1801 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     1861 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     1921 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     1981 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2041 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2101 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2161 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2221 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2281 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2341 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2401 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2461 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     2521 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2581 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2641 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2701 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2761 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2821 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2881 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2941 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3001 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3061 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3121 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3181 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3241 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     3301 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3361 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     3421 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     3481 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     3541 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
     3601 ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta
     3661 ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga
     3721 tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg
     3781 taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc
     3841 agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg
     3901 ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg
     3961 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca
     4021 ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgagatg cgccgcgtgc
     4081 ggctgctgga gatggcggac gcgatggata tgttctgcca agggttggtt tgcgcattca
     4141 cagttctccg caagaattga ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt
     4201 gccgccggct tccattcagg tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg
     4261 gaggcagaca aggtataggg cggcgcctac aatccatgcc aacccgttcc atgtgctcgc
     4321 cgaggcggca taaatcgccg tgacgatcag cggtccagtg atcgaagtta ggctggtaag
     4381 agccgcgagc gatccttgaa gctgtccctg atggtcgtca tctacctgcc tggacagcat
     4441 ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa tggggaaggc
     4501 catccagcct cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc
     4561 ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg
     4621 agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca
     4681 gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg
     4741 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa
     4801 ggagctgact gggttgaagg ctctcaaggg catcggtcga cgctctccct tatgcgactc
     4861 ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa
     4921 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     4981 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     5041 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     5101 tccggcgtag aggatcc
//
