LOCUS       pCR178                  4813 bp    DNA     circular SYN 03-JUN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4813)
  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..4813
                     /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..1866
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     promoter        complement(1893..1911)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1916..2050
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(2167..2185)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    2230..2246
                     /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(2254..2260,2261..2278,2279..2284))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2299..2320
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2608..3196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(3367..4158,4159..4227))
                     /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(4228..4332)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4358..4813)
                     /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 gcgaggcagt gatcaaggag
     1201 ttcatgcggt tcaaggtgca catggagggc tccatgaacg gccacgagtt cgagatcgag
     1261 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     1321 ggtggccccc tgcccttctc ctgggacatc ctgtcccctc agttcatgta cggctccagg
     1381 gccttcatca agcaccccgc cgacatcccc gactactata agcagtcctt ccccgagggc
     1441 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg ccgtgaccgt gacccaggac
     1501 acctccctgg aggacggcac cctgatctac aaggtgaagc tccgcggcac caacttccct
     1561 cctgacggcc ccgtaatgca gaagaagaca atgggctggg aagcgtccac cgagcggttg
     1621 taccccgagg acggcgtgct gaagggcgac attaagatgg ccctgcgcct gaaggacggc
     1681 ggccgctacc tggcggactt caagaccacc tacaaggcca agaagcccgt gcagatgccc
     1741 ggcgcctaca acgtcgaccg caagttggac atcacctccc acaacgagga ctacaccgtg
     1801 gtggaacagt acgaacgctc cgagggccgc cactccaccg gcggcatgga cgagctgtac
     1861 aagtagaatt caaggcctct cgagcctctc gccctatagt gagtcgtatt acgtagatcc
     1921 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     1981 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2041 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     2101 ggaggttttt taattcgcgg cgcgccgcgg cgccaatgca ttgggcccgg tacccagctt
     2161 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     2221 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg
     2281 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2341 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2401 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2461 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2521 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2581 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2641 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2701 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2761 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2821 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2881 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2941 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3001 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     3061 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3121 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3181 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3241 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3301 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3361 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3421 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3481 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3541 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3601 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3661 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3721 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3781 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3841 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3901 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3961 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4021 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4081 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4141 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4201 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4261 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4321 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     4381 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     4441 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     4501 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     4561 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     4621 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     4681 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     4741 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     4801 ctacagggcg cgt
//
