LOCUS       sequence-371216-        7025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses miRFP2 in worm cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7025)
  AUTHORS   Babakhanova S, Jung EE, Namikawa K, Zhang H, Wang Y, Subach OM, 
            Korzhenevskiy DA, Rakitina TV, Xiao X, Wang W, Shi J, Drobizhev M, 
            Park D, Eisenhard L, Tang H, Koster RW, Subach FV, Boyden ES, 
            Piatkevich KD
  TITLE     Rapid directed molecular evolution of fluorescent proteins in 
            mammalian cells.
  JOURNAL   Protein Sci. 2022 Mar;31(3):728-751. doi: 10.1002/pro.4261. Epub 
            2021 Dec 30.
  PUBMED    34913537
REFERENCE   2  (bases 1 to 7025)
  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: "10.1002/pro.4261"; 
            journalName: "Protein Sci"; date: "2022-03"; volume: "31"; issue: 
            "3"; pages: "728-751"
COMMENT     Sequence Label: pSF11-wmiRFP2
FEATURES             Location/Qualifiers
     source          1..7025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4595..4617)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4717..4736
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4818..4836)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4904..5008
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5009..5077,5078..5869)
                     /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     complement(5227..5246)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6040..6628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6529..6548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6782..6799
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6916..6937
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6952..6957,6958..6975,6976..6982)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6990..7006
                     /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     6995..7017
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7014..7025,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7014..7025,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcg cggccgcact gactgggccg gccggatcca
       61 taggtatgcc tgtcattctc tatttttaaa cgttcctgca gcagtagcac aacagagcca
      121 gcagttttcc tcttcttctc catcttcttc tgcttcttcc tctcttccca aaaaaaaaaa
      181 aaaaaaaaaa gtgagggggc tcactgcccc cccccccctc ccccccccct cttctccttg
      241 aagctcatcc agacgtcgct gtcctcctcc tcctcgtccc cggggcgctg tgtgtgtgtg
      301 tgtgtgctcc gtgagcatgt agcagcttca gcccatttaa aggcgcacac tctctgcatc
      361 tctgcgtgtc tcgacactcg ttttttctct tcttttctgc tgcacacaca cacacacaca
      421 cacacacgct catgccagag cacttcttgc tccggggggc tccgcgcgga gcccgagagg
      481 acctcctcct cctttttctt cttcgctttt tagttcttct tgcctcggtc tcgccccttg
      541 ttttgctctc catgtagtat atacatctgg gtgtgtgtgt gtgtgtgcac ctttggagca
      601 ttttttgaac ctttccagaa ctttggagca acattttcag cttgttcagt gcacttttta
      661 ggctgaaaat cctattttta ggttttcgaa agtttttttg gagcattttt cgtgcaattt
      721 aggtgcaaaa ttaaaaaaaa gggttttgta gtgaattaaa atttttttga aactttcgcc
      781 tttttaaaaa aatgtttcta atatctttaa aaattggaat attttggaaa acaatttgtt
      841 ttcaattggc aatttattaa ttctgagctt ctgagcgtaa attttttttt attttggagc
      901 aattctgaga tgtcttagtg catttttcgg gcatacataa aaaaaaattt atcttaataa
      961 taaaaacctt aaatttaggc ttcaccgtct ggaaattcaa ttttttcagc tgaaaaactt
     1021 tcaaaaaaaa aaaaattaaa ttttcacatt tattttctct ggaattttgg agcaatttca
     1081 aagctgtctt agtgcatttt tagtacacaa atattttttc atttttctta aattttaata
     1141 atctaagctt aaataaaggc ttcaccgtct gaaaatgcaa cttttttcag atgaaatatt
     1201 aaaaaaaaaa cctcttttat taattttgtg ctccaaactt tcccacggta cgcaaatcat
     1261 gtaatatgta cttttaaaca ttatttttcc aatcaaaaat atccaacagc ttttccttac
     1321 aatttgaaaa tttttattaa aatttcaaaa aaattttttt tttttttaat ttcgaatttt
     1381 tttaaacttg ttgtaacttt tttccctgtc ggcgccaatt aacaaaatac gtagaatttt
     1441 gcgcttattg ttttgttagt taacaaattt ttgacaactt tcttatcagc tgagatttag
     1501 gtgtttcagt tcgcaatctg cgtctctcct tgctctcctc tattttaaaa cccccataac
     1561 tttgttcttg tgagagttat caaaaaattg tcaactaaca aaatatagag cacaaaaata
     1621 tgcacatttt gtatgttgac aattttttga tacctttttt agttttcgag atatcttagt
     1681 tttagtttag ctgtaccttt ttctggcaaa aaacttccaa atttcggcaa tttttcggca
     1741 aatttcccgg caaatttcgc caagtacatc gggctctcag acaagtggtc tgtgagtgga
     1801 agtcgataaa agtagtttac gtgctctccg tccccccgct tcttctccgt ctctgcttct
     1861 atttctccat tgactagtcc cactactttt tacggggcgg agggctctct tgcactccag
     1921 cttcttctct ttttgtgctc ttttttgccc gtatgtgggt gctcatttat ttttctttgg
     1981 ttttaaaata aattcggttt attaggctac tcgtaaaaat gaagaaaaaa cttcaaattc
     2041 aaacgacatt aaaaaaaaat ttaataaaaa ttttttgaat tgctcaaaag ccccacctat
     2101 atatatatat agctccgcct acttctcgca cagaccccgc ccactttttg actccgcccc
     2161 cttattggca aaagttcttg aaaaaaataa aaataaaaat tcaaattgaa acaatttttt
     2221 tttaatgaag tagttttttt ctaaattgat gaaaatttct aaacagttta caagcaccgc
     2281 ccactttttg actccgcccc tttgttggaa aaattcttgt aaaaaaaaat aaaaaataaa
     2341 agcttctctt tcaaacgatt tttagttggt ctttgtatga aaagtttttt tcctaaaata
     2401 gaaatgtctg aataatttgc aagctccgcc cttttcttac ccagactccg cccctttatt
     2461 gactaatttt tttttgtcca tttacctgaa gttcaaaaaa aaaatttaca gtttcataga
     2521 gtaataattt ttcaaaaatc tctttttcca atgtttaatg tgtcattata aacccgccca
     2581 gttttaagct ccgcccattt aggccccgcc catttatgac ccaaatattt ttttctctat
     2641 tcacccgtgg atttccctcc aacaagttac cccaaaaata aaaataaaaa ttcctgcagg
     2701 cgcccaccgg gatccggcgc gcctctagag gatccccggg attggccaaa ggacccaaag
     2761 gtatgtttcg aatgatacta acataacata gaacattttc aggaggaccc ttggctagcg
     2821 tcgacggtac catggtagcc ggacacgcct caggtagtcc agacttcggc accgcctcgc
     2881 caagtgactg cgagcatgag gagattcatc ttgctggctc tatccaacca cacggcactc
     2941 tgttggtcgt gtcggaacct gaccatagaa tcattcaagc atccgcgaac gcggctgaat
     3001 ttttaaacct gggttccgta ttgggtatac cattagcgga aatagacggc gacctcctca
     3061 taaaaatctt gcctcacctt gacctcaccg cagaaggcat gccggtcgca gttcgttgcc
     3121 gaatcggaaa tccttcgaca gagtatgatg gccttatgca ccgacctccg gagggcggtc
     3181 tgattataga gctggagaga gcaggcccat ctatagacct ttccggaaca cttgcaccgg
     3241 cgctcgaacg aataagaacg gcaggcagtt tgcgtgcatt gtgcgatgac acagtcttac
     3301 tctttcagca atgcacaggt tatgaccgag tcatggtgta tcgtttcgat gaaagaggtc
     3361 atggcgaggt ttactcggag atccatgtga ccggactcga atcatatttt ggcaatcgtt
     3421 acccgagttc gactgtaccg caaatggcaa gaagattata tgtacgtcaa cgtgtgcgag
     3481 tgttagttga tgtaacttac cagccagtcc cgttggagcc gcgactctca ccattaacgg
     3541 gcagagatct ggatatgtct ggatgcttct tacgttcgat gtcaccatgt catctgcagt
     3601 tcctgaaaaa catgggcgtt agagccactc tggttgtatc gctcgttgta ggaggcaaac
     3661 tttggggact ggtaatatgc catcattacc ttcctcgatt catacatttc gaattacgag
     3721 ctatttgcgt tctgctggct gaagcaatag ctacacgaat caccgccttg ggttaagagc
     3781 tccgcatcgg ccgctgtcat cagatcgcca tctcgcgccc gtgcctctga cttctaagtc
     3841 caattactct tcaacatccc tacatgctct ttctccctgt gctcccaccc cctatttttg
     3901 ttattatcaa aaaacttctc ttaatttctt tgttttttag cttcttttaa gtcacctcta
     3961 acaatgaaat tgtgtagatt caaaaataga attaattcgt aataaaaagt cgaaaaaaat
     4021 tgtgctccct ccccccatta ataataattc tatcccaaaa tctacacaat gttctgtgta
     4081 cacttcttat gttttttact tctgataaat ttttttgaaa catcatagaa aaaaccgcac
     4141 acaaaatacc ttatcatatg ttacgtttca gtttatgacc gcaattttta tttcttcgca
     4201 cgtctgggcc tctcatgacg tcaaatcatg ctcatcgtga aaaagttttg gagtattttt
     4261 ggaatttttc aatcaagtga aagtttatga aattaatttt cctgcttttg ctttttgggg
     4321 tttcccctat tgtttgtcaa gatttcgagg acggcgtttt tcttgctaaa atcacaagta
     4381 ttgatgagca cgatgcaaga aagatcggaa gaaggtttgg gtttgaggct cagtggaagg
     4441 tgagtagaag ttgataattt gaaagtggag tagtgtctat ggggtttttg ccttaaatga
     4501 cagaatacat tcccaatata ccaaacataa ctgtttctac tagtcggccg tacgggccct
     4561 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     4621 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     4681 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     4741 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     4801 ggcggcctta attaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4861 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4921 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4981 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     5041 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5101 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5161 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5221 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5281 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5341 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5401 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5461 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5521 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5581 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5641 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5701 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5761 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5821 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5881 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5941 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6001 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6061 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6121 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6181 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6241 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6301 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6361 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6421 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6481 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6541 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6601 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6661 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6721 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6781 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     6841 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     6901 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     6961 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     7021 cagct
//
