LOCUS       sequence-335108-        3729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Strong mCherry fluorescence from Bxb1 landing pad.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3729)
  AUTHORS   Shukla N, Roelle SM, Suzart VG, Bruchez AM, Matreyek KA
  TITLE     Mutants of human ACE2 differentially promote SARS-CoV and SARS-CoV-2
            spike mediated infection.
  JOURNAL   PLoS Pathog. 2021 Jul 16;17(7):e1009715. doi: 
            10.1371/journal.ppat.1009715. eCollection 2021 Jul.
  PUBMED    34270613
REFERENCE   2  (bases 1 to 3729)
  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: "PLoS 
            Pathog."; date: "2021-07-16"; volume: "17(7):e1009715. doi"; pages: 
            " 10.1371/journal.ppat.1009715. eCollection 2021 Jul"
COMMENT     Sequence Label: AttB_mCherry
FEATURES             Location/Qualifiers
     source          1..3729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     113..130
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    247..268
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(283..288,289..306,307..313)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    321..337
                     /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     326..348
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     345..361
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     345..361
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        382..400
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    409..530
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(420..439)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     474..493
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(644..1354)
                     /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(922..941)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1191..1209
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1351..1360
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1385..1422
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     primer_bind     complement(1495..1514)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1496..1514)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1521..1538)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1521..1537)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1530..1552)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1679..2134
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1766..1785)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1976..1997
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2160..2264
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2265..2333,2334..3125)
                     /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"
     primer_bind     complement(2483..2502)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
       61 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagt gcagcgagtc
      121 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
      181 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
      241 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
      301 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
      361 ccatgattac gccaagctcg aaattaaccc tcactaaagg ggatctccta agatacattg
      421 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
      481 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
      541 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
      601 aaaacctcta caaatgtggt atggctgatt atgatcagga tccctacttg tacagctcgt
      661 ccatgccgcc ggtggagtgg cggccctcgg cgcgttcgta ctgttccacg atggtgtagt
      721 cctcgttgtg ggaggtgatg tccaacttga tgttgacgtt gtaggcgccg ggcagctgca
      781 cgggcttctt ggccttgtag gtggtcttga cctcagcgtc gtagtggccg ccgtccttca
      841 gcttcagcct ctgcttgatc tcgcccttca gggcgccgtc ctcggggtac atccgctcgg
      901 aggaggcctc ccagcccatg gtcttcttct gcattacggg gccgtcggag gggaagttgg
      961 tgccgcgcag cttcaccttg tagatgaact cgccgtcctg cagggaggag tcctgggtca
     1021 cggtcaccac gccgccgtcc tcgaagttca tcacgcgctc ccacttgaag ccctcgggga
     1081 aggacagctt caagtagtcg gggatgtcgg cggggtgctt cacgtaggcc ttggagccgt
     1141 acatgaactg aggggacagg atgtcccagg cgaagggcag ggggccaccc ttggtcacct
     1201 tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc gccctcgccc tcgatctcga
     1261 actcgtggcc gttcacggag ccctccatgt gcaccttgaa gcgcatgaac tccttgatga
     1321 tggccatgtt atcctcctcg cccttgctca ccatggtggc gtgtagttgc ggtataatct
     1381 gcggatgatc ctgacgacgg agaccgccgt cgtcgacaag ccggctcgag cgctgagtcc
     1441 ggaatgccgt gacctacatc gtcgacctcg agggggggcc cggtacccaa ttcgccctat
     1501 agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     1561 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     1621 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     1681 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1741 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1801 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1861 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1921 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1981 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2041 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2101 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     2161 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2221 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2281 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2341 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2401 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2461 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2521 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2581 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2641 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2701 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2761 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     2821 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2881 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2941 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3001 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3061 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3121 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3181 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3241 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3301 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3361 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3421 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3481 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3541 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3601 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3661 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3721 aggcggaca
//
