LOCUS       sequence-67925-e        5068 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 5068)
  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 5068)
  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-hCaM
FEATURES             Location/Qualifiers
     source          1..5068
                     /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"
     CDS             278..727
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="MADQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPT
                     EAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAA
                     ELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     complement(803..820)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      973..1059
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1151..1178
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1198..1289
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1290..1358,1359..2150)
                     /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(1508..1527)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2321..2909
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2810..2829
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3063..3080
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3095..3235
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3161..3180)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3321..3343
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3337..3528)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4986..5005
                     /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 gctgaccagc tgactgagga
      301 gcagattgca gagttcaagg aggccttctc cctctttgac aaggatggag atggcactat
      361 caccaccaag gagttgggga cagtgatgag atccctggga cagaacccca ctgaagcaga
      421 gctgcaggat atgatcaatg aggtggatgc agatgggaac gggaccattg acttcccgga
      481 gttcctgacc atgatggcca gaaagatgaa ggacacagac agtgaggagg agatccgaga
      541 ggcgttccgt gtctttgaca aggatgggaa tggctacatc agcgccgcag agctgcgtca
      601 cgtaatgacg aacctggggg agaagctgac cgatgaggag gtggatgaga tgatcaggga
      661 ggctgacatc gatggagatg gccaggtcaa ttatgaagag tttgtacaga tgatgactgc
      721 aaagtgaagg ccccccggga tcctctagag tcgacctgca gcccaagctt ggctgttttg
      781 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga
      841 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact
      901 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga
      961 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     1021 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     1081 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat
     1141 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttt
     1201 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     1261 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     1321 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     1381 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     1441 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     1501 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     1561 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     1621 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     1681 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     1741 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     1801 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     1861 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     1921 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     1981 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     2041 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     2101 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     2161 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     2221 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     2281 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2341 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2401 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2461 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2521 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2581 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2641 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2701 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2761 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2821 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2881 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2941 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3001 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3061 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     3121 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
     3181 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     3241 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     3301 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     3361 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     3421 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     3481 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     3541 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     3601 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     3661 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     3721 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
     3781 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
     3841 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     3901 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     3961 cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat
     4021 gcgccgcgtg cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt
     4081 ttgcgcattc acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc
     4141 gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga
     4201 cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc
     4261 catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca gcggtccagt gatcgaagtt
     4321 aggctggtaa gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc
     4381 ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata
     4441 atggggaagg ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg
     4501 gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg
     4561 acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc
     4621 gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt
     4681 cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc
     4741 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg acgctctccc
     4801 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
     4861 gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct
     4921 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
     4981 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
     5041 gccacgatgc gtccggcgta gaggatcc
//
