LOCUS       pCR175                  4834 bp    DNA     circular SYN 01-JUN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4834)
  AUTHORS   Christian Renicke
  TITLE     Direct Submission
  JOURNAL   Exported Jun 3, 2025 from SnapGene 8.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        78..1062
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1097..1115
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             1168..1886
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        complement(1914..1932)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1937..2071
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(2188..2206)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    2251..2267
                     /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)."
     promoter        complement(join(2275..2281,2282..2299,2300..2305))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2320..2341
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2629..3217)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(3388..4179,4180..4248))
                     /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"
     promoter        complement(4249..4353)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4379..4834)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gtcgatcgac catagccaat tcaatatggc gtatatggac tcatgccaat
      121 tcaatatggt ggatctggac ctgtgccaat tcaatatggc gtatatggac tcgtgccaat
      181 tcaatatggt ggatctggac cccagccaat tcaatatggc ggacttggca ccatgccaat
      241 tcaatatggc ggacttggca ctgtgccaac tggggagggg tctacttggc acggtgccaa
      301 gtttgaggag gggtcttggc cctgtgccaa gtccgccata ttgaattggc atggtgccaa
      361 taatggcggc catattggct atatgccagg atcaatatat aggcaatatc caatatggcc
      421 ctatgccaat atggctattg gccaggttca atactatgta ttggccctat gccatatagt
      481 attccatata tgggttttcc tattgacgta gatagcccct cccaatgggc ggtcccatat
      541 accatatatg gggcttccta ataccgccca tagccactcc cccattgacg tcaatggtct
      601 ctatatatgg tctttcctat tgacgtcata tgggcggtcc tattgacgta tatggcgcct
      661 cccccattga cgtcaattac ggtaaatggc ccgcctggct caatgcccat tgacgtcaat
      721 aggaccaccc accattgacg tcaatgggat ggctcattgc ccattcatat ccgttctcac
      781 gccccctatt gacgtcaatg acggtaaatg gcccacttgg cagtacatca atatctatta
      841 atagtaactt ggcaagtaca ttactattgg aagtacgcca gggtacattg gcagtactcc
      901 cattgacgtc aatggcggta aatggcccgc gatggctgcc aagtacatcc ccattgacgt
      961 caatggggag gggcaatgac gcaaatgggc gttccattga cgtaaatggg cggtaggcgt
     1021 gcctaatggg aggtctatat aagcaatgct cgtttaggga accgccattc tgcctgggga
     1081 cgtcggagca agcttgattt aggtgacact atagaataca agctacttgt tctttttgca
     1141 ggatcccatc gattcgaatt cgtcgacatg gtgagcaagg gcgaggagct gttcaccggg
     1201 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1261 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     1321 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgtcctgggg cgtgcagtgc
     1381 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1441 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1501 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1561 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actactttag cgacaacgtc
     1621 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     1681 atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1741 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac
     1801 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     1861 ctcggcatgg acgagctgta caagtagaat tcaaggcctc tcgagcctct cgccctatag
     1921 tgagtcgtat tacgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     1981 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     2041 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     2101 tcaggttcag ggggaggtgt gggaggtttt ttaattcgcg gcgcgccgcg gcgccaatgc
     2161 attgggcccg gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     2221 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     2281 acgagccggg agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     2341 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     2401 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     2461 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2521 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2581 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2641 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2701 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2761 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2821 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2881 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2941 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3001 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3061 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3121 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3181 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3241 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3301 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3361 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3421 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3481 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3541 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3601 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3661 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3721 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3781 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3841 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3901 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3961 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4021 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4081 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4141 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4201 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4261 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4321 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     4381 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     4441 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     4501 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     4561 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     4621 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     4681 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     4741 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     4801 cccgccgcgc ttaatgcgcc gctacagggc gcgt
//
