LOCUS       sequence-330922-        3393 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB5r/attB1r entry clone for generation of N-terminal mTFP1
            fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3393)
  AUTHORS   Wall VE, Garvey LA, Mehalko JL, Procter LV, Esposito D
  TITLE     Combinatorial assembly of clone libraries using site-specific 
            recombination.
  JOURNAL   Methods Mol Biol. 2014;1116:193-208. doi: 
            10.1007/978-1-62703-764-8_14.
  PUBMED    24395366
REFERENCE   2  (bases 1 to 3393)
  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: "Methods Mol
            Biol."; date: "2014"; volume: "1116:193-208. doi"; pages: " 
            10.1007/978-1-62703-764-8_14"
COMMENT     Sequence Label: C512-E11: attB5r-mTFP1-nostop-attB1r
FEATURES             Location/Qualifiers
     source          1..3393
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(44..167)
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             171..878
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     protein_bind    879..1003
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1075..1094)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1076..1094)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1099..1115)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1099..1115)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1540..2331
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      2424..3012
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2913..2932
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3147..3174)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3266..3352)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agctcgaatt ctacagtcac taataccatc taagtagttg attcatagtg actgcatatg
       61 ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat
      121 atttatatca ttttacgttt ctcgttcaac ttttgtatac aaagtggacc atggtgagca
      181 agggcgagga gaccacaatg ggcgtaatca agcccgacat gaagatcaag ctgaagatgg
      241 agggcaacgt gaatggccac gccttcgtga tcgagggcga gggcgagggc aagccctacg
      301 acggcaccaa caccatcaac ctggaggtga aggagggagc ccccctgccc ttctcctacg
      361 acattctgac caccgcgttc gcctacggca acagggcctt caccaagtac cccgacgaca
      421 tccccaacta cttcaagcag tccttccccg agggctactc ttgggagcgc accatgacct
      481 tcgaggacaa gggcatcgtg aaggtgaagt ccgacatctc catggaggag gactccttca
      541 tctacgagat acacctcaag ggcgagaact tcccccccaa cggccccgtg atgcagaaga
      601 agaccaccgg ctgggacgcc tccaccgaga ggatgtacgt gcgcgacggc gtgctgaagg
      661 gcgacgtcaa gcacaagctg ctgctggagg gcggcggcca ccaccgcgtt gacttcaaga
      721 ccatctacag ggccaagaag gcggtgaagc tgcccgacta tcactttgtg gaccaccgca
      781 tcgagatcct gaaccacgac aaggactaca acaaggtgac cgtttacgag agcgccgtgg
      841 cccgcaactc caccgacggc atggacgagc tgtacaagac aactttgtac aaaaaagttg
      901 aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg cataaaaaac
      961 agactacata atactgtaaa acacaacata tgcagtcact atgaaccaac tacttagatg
     1021 gtattagtga cctgtagaat tcgagctcta gagctgcagg gcggccgcga tatcccctat
     1081 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     1141 ctctgatgtt acattgcaca agatcggtac ccggggatcc tctagaccag ccaggacaga
     1201 aatgcctcga cttcgctgct acccaaggtt gccgggtgac gcacaccgtg gaaacggatg
     1261 aaggcacgaa cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt
     1321 atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg
     1381 gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc
     1441 aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt
     1501 acgcagcagg gcagtcgccc taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc
     1561 gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg
     1621 ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat
     1681 attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc
     1741 aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa
     1801 gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg
     1861 caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc
     1921 gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt
     1981 ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat
     2041 gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg
     2101 cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc
     2161 gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct
     2221 agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa
     2281 gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata accctcgagc
     2341 cacccatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     2401 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2461 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2521 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     2581 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2641 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2701 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2761 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2821 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2881 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2941 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3001 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3061 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgct
     3121 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
     3181 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca
     3241 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
     3301 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
     3361 cagttcccta ctctcgcgtt aacgctacca tgg
//
