LOCUS       sequence-394836-        3925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Strong mCherry fluorescence recombination plasmid for Matreyek 
            Bxb1(GA) landing pad.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3925)
  AUTHORS   Roelle SM, Kamath ND, Matreyek KA
  TITLE     Mammalian Genomic Manipulation with Orthogonal Bxb1 DNA Recombinase 
            Sites for the Functional Characterization of Protein Variants.
  JOURNAL   ACS Synth Biol. 2023 Nov 3. doi: 10.1021/acssynbio.3c00355.
  PUBMED    37922210
REFERENCE   2  (bases 1 to 3925)
  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: "ACS Synth 
            Biol. 2023 Nov 3. doi: 10.1021/acssynbio.3c00355."
COMMENT     Sequence Label: G421C_BattB(GA)_mCherry
FEATURES             Location/Qualifiers
     source          1..3925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     620..639
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(858..962)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(988..1443)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1125..1146)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1337..1356
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1570..1592
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1584..1601
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1585..1601
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1608..1627
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1608..1626
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1700..1737
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     regulatory      1762..1771
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1768..2478
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1913..1931)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2181..2200
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2592..2713
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2629..2648)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2683..2702
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(2722..2740)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2761..2777)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2761..2777)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2774..2796)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2785..2801
                     /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(2809..2815,2816..2833,2834..2839))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2854..2875
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2992..3009)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3163..3751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3243..3262)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3922..3925,1..788,789..857))
                     /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"
ORIGIN
        1 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
       61 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
      121 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
      181 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
      241 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
      301 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
      361 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
      421 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
      481 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
      541 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
      601 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
      661 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
      721 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
      781 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
      841 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
      901 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat agtggttccg
      961 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     1021 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     1081 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     1141 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     1201 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     1261 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     1321 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     1381 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     1441 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     1501 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     1561 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa tacgactcac
     1621 tatagggcga attgggtacc gggccccccc tcgaggtcga cgatgtaggt cacggcattc
     1681 cggactcagc gctcgagccg gcttgtcgac gacggcggac tccgtcgtca ggatcatccg
     1741 cagattatac cgcaactaca cgccaccatg gtgagcaagg gcgaggagga taacatggcc
     1801 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     1861 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     1921 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     1981 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     2041 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     2101 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     2161 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     2221 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     2281 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     2341 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     2401 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     2461 gacgagctgt acaagtaggg atcctgatca taatcagcca taccacattt gtagaggttt
     2521 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     2581 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     2641 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     2701 tcaatgtatc ttaggagatc ccctttagtg agggttaatt tcgagcttgg cgtaatcatg
     2761 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     2821 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     2881 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     2941 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3001 tgactcgctg cactcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3061 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3121 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3181 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3241 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3301 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3361 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3421 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3481 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3541 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3601 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3661 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3721 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3781 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3841 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3901 tgagtaaact tggtctgaca gttac
//
