LOCUS       sequence-9475-ex        3388 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 3388)
  TITLE     FQG plasmids
REFERENCE   2  (bases 1 to 3388)
  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: pEF 3-CFP-2
FEATURES             Location/Qualifiers
     source          1..3388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(119..138)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(120..138)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(143..159)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(143..159)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(261..280)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             272..1081
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(339..358)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1228..1816
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1717..1736
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1970..1987
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2146..2173)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2265..2351)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2400..2422
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2414..2431
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2415..2431
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2448..2543
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(2558..2560,2561..3271)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2600..2621)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2861..2880)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3208..3229
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3272..3337
                     /label=MCS
                     /note="multiple cloning site"
     CDS             3350..3376
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(join(3378..3388,1..89))
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc
       61 actatcagtc aaaataaaat cattatttgc catccagctg cagggcggcc gcgatatccc
      121 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
      181 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
      241 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
      301 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
      361 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
      421 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
      481 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
      541 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
      601 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
      661 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
      721 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
      781 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
      841 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
      901 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
      961 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     1021 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     1081 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     1141 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     1201 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1261 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1321 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1381 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1441 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1501 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1561 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1621 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1681 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1741 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1801 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1861 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1921 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     1981 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     2041 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     2101 aattaatacg cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc
     2161 aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac
     2221 cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg
     2281 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt
     2341 cgttttattt gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc
     2401 ccagtcacga cgttgtaaaa cgacggccag tcttaagctc gggccccaaa taatgatttt
     2461 attttgactg atagtgacct gttcgttgca acaaattgat gagcaatgct tttttataat
     2521 gccaactttg tataataaag ttgctgtggt gcctagggtg agcaagggcg aggagctgtt
     2581 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     2641 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     2701 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctggggcgt
     2761 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     2821 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     2881 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     2941 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acatcagcca
     3001 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg
     3061 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     3121 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     3181 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     3241 gatcactctc ggcatggacg agctgtacaa gtccggactc agatctcgag ctcaagcttc
     3301 gaattctgca gtcgacggta ccgcgggccc gggatccacc ggatctagat acccatacga
     3361 tgttccagat tacgcttacc cagctttc
//
