LOCUS       sequence-199613-        3289 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  create your own 2in1 constructs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3289)
  AUTHORS   Grefen C, Blatt MR
  TITLE     A 2in1 cloning system enables ratiometric bimolecular fluorescence 
            complementation (rBiFC).
  JOURNAL   Biotechniques. 2012 Oct 2;53(5):311-14. doi: 10.2144/000113941.
  PUBMED    23066669
REFERENCE   2  (bases 1 to 3289)
  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"; doi: "10"; journalName: 
            "Biotechniques"; date: "2012-10-2- 2"; volume: "53"; issue: "5"; 
            pages: "311-14"
COMMENT     Sequence Label: pUC57-R3R2
FEATURES             Location/Qualifiers
     source          1..3289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    415..538
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(573..578,579..596,597..603)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    611..627
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     635..651
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(912..1036)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1068..1084)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1092..1108
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1116..1122,1123..1140,1141..1146))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1161..1182
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1299..1316)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1470..2058)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1550..1569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2229..3020,3021..3089))
                     /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     2852..2871
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3090..3194)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3262..3280
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cactagtcaa ttggcaactt
      421 tgtataataa agttgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga
      481 ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag tcactatggc
      541 ggccgcgggc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
      601 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
      661 cccgtccctg cacgcctgtc gttcaaccct ggaagatccg cgtgtcccga gttctaacag
      721 tctggccgtc gttctgcagc gtcgcgattg ggaaaacccg ggcgtgaccc aactgaatcg
      781 tctggcggcc catccgccgt ttgcgtcgtg gcgtaactca gaagaagccc gtaccgaccg
      841 tccgtctcaa caactgcgct cctaacacta taaaaagctg atgcagacgg ggcttttttc
      901 tgcaggtcga ccatagtgac tggatatgtt gtgttttaca gtattatgta gtctgttttt
      961 tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct cgttcagctt
     1021 tcttgtacaa agtggtgtct aggttataaa agcttggcgt aatcatggtc atagctgttt
     1081 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     1141 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     1201 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     1261 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     1321 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     1381 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1441 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1501 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1561 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1621 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1681 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1741 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1801 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1861 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     1921 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     1981 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2041 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2101 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2161 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2221 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     2281 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     2341 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     2401 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     2461 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     2521 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     2581 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     2641 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     2701 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     2761 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     2821 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     2881 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     2941 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3001 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     3061 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3121 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3181 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     3241 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc
//
