LOCUS       sequence-347642-        3529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB2r/attB3 entry clone for construction of C-terminal 
            mTFP1 fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3529)
  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 3529)
  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: C232-E08: attB2r-mTFP1-attB3
FEATURES             Location/Qualifiers
     source          1..3529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     95..117
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     109..126
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     110..126
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(224..347)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             353..1060
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="VSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDGT
                     NTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTFE
                     DKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLKG
                     DVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESAV
                     ARNSTDGMDELYK"
     protein_bind    complement(1062..1157)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1568..2359
                     /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      2452..3040
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2941..2960
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3194..3211
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3370..3397)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(join(3489..3529,1..46))
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc
       61 cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg taaaacgacg
      121 gccagtctta aaattcgagc tcggtccgat atccttaagc tcgggccctg cagctctaga
      181 gctcgaattc tacaggtcac taataccatc taagtagttg gttcatagtg actgcatatg
      241 ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat
      301 atttatatca ttttacgttt ctcgttcaac tttcttgtac aaagtgggcg gcgtgagcaa
      361 gggcgaggag accacaatgg gcgtaatcaa gcccgacatg aagatcaagc tgaagatgga
      421 gggcaacgtg aatggccacg ccttcgtgat cgagggcgag ggcgagggca agccctacga
      481 cggcaccaac accatcaacc tggaggtgaa ggagggagcc cccctgccct tctcctacga
      541 cattctgacc accgcgttcg cctacggcaa cagggccttc accaagtacc ccgacgacat
      601 ccccaactac ttcaagcagt ccttccccga gggctactct tgggagcgca ccatgacctt
      661 cgaggacaag ggcatcgtga aggtgaagtc cgacatctcc atggaggagg actccttcat
      721 ctacgagata cacctcaagg gcgagaactt cccccccaac ggccccgtga tgcagaagaa
      781 gaccaccggc tgggacgcct ccaccgagag gatgtacgtg cgcgacggcg tgctgaaggg
      841 cgacgtcaag cacaagctgc tgctggaggg cggcggccac caccgcgttg acttcaagac
      901 catctacagg gccaagaagg cggtgaagct gcccgactat cactttgtgg accaccgcat
      961 cgagatcctg aaccacgaca aggactacaa caaggtgacc gtttacgaga gcgccgtggc
     1021 ccgcaactcc accgacggca tggacgagct gtacaagtaa tcaactttat tatacaaagt
     1081 tggcattata aaaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     1141 aaataaaatc attatttgga gctccatggt agcgttaacg cggccgcgat atcggtaccc
     1201 ggggatcctc tagaccagcc aggacagaaa tgcctcgact tcgctgctac ccaaggttgc
     1261 cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat aagcctgttc
     1321 ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac
     1381 cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt
     1441 tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg
     1501 tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt
     1561 aaacattatg agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg
     1621 cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt
     1681 ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct
     1741 tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg
     1801 agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc
     1861 gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct
     1921 tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc
     1981 aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc
     2041 tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga
     2101 ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt
     2161 aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc
     2221 ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg
     2281 cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac
     2341 caaggtagtc ggcaaataac cctcgagcca cccatgacca aaatccctta acgtgagtta
     2401 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2461 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2521 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2581 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     2641 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2701 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     2761 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     2821 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2881 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2941 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3001 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3061 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     3121 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     3181 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     3241 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     3301 gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt
     3361 agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt
     3421 tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct
     3481 cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacaga
//
