LOCUS       sequence-334120-        2785 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of mClover3 green fluorescent protein .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2785)
  TITLE     Verstreken lab plasmids
REFERENCE   2  (bases 1 to 2785)
  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: pIVTR(T7G)-mClover3
FEATURES             Location/Qualifiers
     source          1..2785
                     /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"
     regulatory      77..86
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(83..85,86..88,89..802)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(128..149)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     736..757
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     rep_origin      complement(1030..1618)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1110..1129)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1789..2580,2581..2649))
                     /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     2412..2431
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2650..2754)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taatacgact cactataggg acatttgctt ctgacacaac tgtgttcact agcaacctca
       61 aacagaattc gagctcgcca ccatggtgtc taagggcgag gagctgttca ccggggtggt
      121 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tccgcggcga
      181 gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa
      241 gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcgtgg cctgcttcag
      301 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
      361 cgtccaggag cgcaccatct ctttcaagga cgacggtacc tacaagaccc gcgccgaggt
      421 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
      481 ggacggcaac atcctggggc acaagctgga gtacaacttc aacagccact acgtctatat
      541 cacggccgac aagcagaaga actgcatcaa ggctaacttc aagatccgcc acaacgttga
      601 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
      661 cgtgctgctg cccgacaacc actacctgag ccatcagtcc aagctgagca aagaccccaa
      721 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga ttacacatgg
      781 catggacgag ctgtacaagt aatctagagt cgacctgcag gcatgcaagc ttgctcgctt
      841 tcttgctgtc caatttctat taaaggttcc tttgttccct aagtccaact actaaactgg
      901 gggatattat gaagggcctt gagcatctgg attctgccta ataaaaaaca tttattttca
      961 ttgcaaaacc atgggagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1021 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1081 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1141 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1201 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1261 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1321 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1381 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1441 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1501 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1561 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1621 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1681 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1741 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     1801 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     1861 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     1921 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     1981 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     2041 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     2101 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     2161 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     2221 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     2281 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     2341 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     2401 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     2461 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     2521 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     2581 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     2641 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     2701 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     2761 tccccgaaaa gtgccacctg acgtc
//
