LOCUS       sequence-211954-        5469 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 5469)
  TITLE     C-terminal fluorescent fusion vectors for E. coli expression
REFERENCE   2  (bases 1 to 5469)
  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"
COMMENT     Sequence Label: pET mCherry LIC cloning vector (u-mCherry)
FEATURES             Location/Qualifiers
     source          1..5469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             108..130
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             171..878
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(313..331)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     581..600
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(999..1017)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1013..1060
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1426..1431,1432..1448,1449..1454))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1481..1499)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1567..1671
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1672..1740,1741..2532)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1890..1909)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2703..3291
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3192..3211
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3445..3462
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3477..3616
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3542..3561)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3702..3724
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3718..3909)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5367..5386
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagtgccggc tccggagagc
       61 tctttaatta agcggccgcc ctgcaggact cgagttctag aaataatttt gtttaacttt
      121 aagaaggaga tatagatatc ctctcatcct ccaacggcag cagcggatcc gtgagcaagg
      181 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
      241 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
      301 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
      361 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
      421 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
      481 aggacggcgg cgtggtgacc gtgacccagg actcctccct ccaggacggc gagttcatct
      541 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
      601 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
      661 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca
      721 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg
      781 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
      841 gccactccac cggcggcatg gacgagctgt acaagtagta accaactcca taaggatccg
      901 cgatcgcggc gcgccacctg gtggccggcc ggtaccacgc gtgcgcgctg atccggctgc
      961 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     1021 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     1081 cggacatcca caggacgggt gtggtcgcca tgatcgcgta gtcgatagtg gctccaagta
     1141 gcgaagcgag caggactggg cggcggccaa agcggtcgga cagtgctccg agaacgggtg
     1201 cgcatagaaa ttgcatcaac gcatatagcg ctagcagcac gccatagtga ctggcgatgc
     1261 tgtcggaatg gacgacatcc cgcaagaggc ccggcagtac cggcataacc aagcctatgc
     1321 ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta gatttcatac
     1381 acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa agcttatcga
     1441 tgataagctg tcaaacatga gaattcttga agacgaaagg gcctcgtgat acgcctattt
     1501 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     1561 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1621 atgagacaat aaccctgata aatgcttcaa taacattgaa aaaggaagag tatgagtatt
     1681 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     1741 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     1801 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     1861 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     1921 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     1981 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2041 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2101 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2161 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     2221 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2281 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2341 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2401 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2461 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2521 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     2581 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     2641 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2701 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2761 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2821 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     2881 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2941 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3001 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3061 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3121 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3181 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3241 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3301 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3361 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3421 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     3481 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca
     3541 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
     3601 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3661 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3721 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     3781 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     3841 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     3901 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
     3961 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
     4021 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
     4081 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca
     4141 tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact
     4201 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca
     4261 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca
     4321 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggcca
     4381 ggacccaacg ctgcccgaga tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga
     4441 tatgttctgc caagggttgg tttgcgcatt cacagttctc cgcaagaatt gattggctcc
     4501 aattcttgga gtggtgaatc cgttagcgag gtgccgccgg cttccattca ggtcgaggtg
     4561 gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga caaggtatag ggcggcgcct
     4621 acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc
     4681 agcggtccaa tgatcgaagt taggctggta agagccgcga gcgatccttg aagctgtccc
     4741 tgatggtcgt catctacctg cctggacagc atggcctgca acgcgggcat cccgatgccg
     4801 ccggaagcga gaagaatcat aatggggaag gccatccagc ctcgcgtcgc gaacgccagc
     4861 aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa
     4921 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc
     4981 gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc
     5041 cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg
     5101 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     5161 ggcatcggtc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     5221 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     5281 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     5341 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     5401 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
     5461 ccgcgaaat
//
