LOCUS       sequence-250841-        4222 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid has a CaMV 35S promoter in a forward sequence orientation 
            and an inverted EGFP coding sequence (reverse complement) flanked by
            attB and attP integrase 13 attachment sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4222)
  AUTHORS   Gomide MS, Sales TT, Barros LRC, Limia CG, de Oliveira MA, 
            Florentino LH, Barros LMG, Robledo ML, Jose GPC, Almeida MSM, Lima 
            RN, Rehen SK, Lacorte C, Melo EO, Murad AM, Bonamino MH, Coelho CM, 
            Rech E
  TITLE     Genetic switches designed for eukaryotic cells and controlled by 
            serine integrases.
  JOURNAL   Commun Biol. 2020 May 22;3(1):255. doi: 10.1038/s42003-020-0971-8.
  PUBMED    32444777
REFERENCE   2  (bases 1 to 4222)
  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: "Commun 
            Biol."; date: "2020-05-22"; volume: "3(1):255. doi"; pages: " 
            10.1038/s42003-020-0971-8"
COMMENT     Sequence Label: INCTbiosyn-p35S-GFP(rc)13
FEATURES             Location/Qualifiers
     source          1..4222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     201..223
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     215..232
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     216..232
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        314..658
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     602..622
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             complement(join(762..1475,1476..1478,1479..1481))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(807..828)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1156..1175
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1415..1436
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     terminator      1556..1809
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1838..1854)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1838..1854)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1851..1873)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1862..1878
                     /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(1886..1892,1893..1910,1911..1916))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1931..1952
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2069..2086)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2240..2828)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2320..2339)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2999..3790,3791..3859))
                     /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     3622..3641
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3860..3964)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4032..4050
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4088..4110)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     join(4210..4222,1..7)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
       61 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
      121 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
      181 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct
      241 tgcatgcctg caggtcgact ctagaggatc ccctaattca tcaaattata actatataga
      301 ccctaatttc atatgagact tttcaacaaa gggtaatatc cggaaacctc ctcggattcc
      361 attgcccagc tatctgtcac tttattgtga agatagtgga aaaggaaggt ggctcctaca
      421 aatgccatca ttgcgataaa ggaaaggcca tcgttgaaga tgcctctgcc gacagtggtc
      481 ccaaagatgg acccccaccc acgaggagca tcgtggaaaa agaagacgtt ccaaccacgt
      541 cttcaaagca agtggattga tgtgatatct ccactgacgt aagggatgac gcacaatccc
      601 actatccttc gcaagaccct tcctctatat aaggaagttc atttcatttg gagaggacta
      661 actgtgccaa attatcttcc aaactcctaa tccaattgca tacattgttg ttgtttttcc
      721 agatccagtt ggtcctgtaa atataagcaa tccatgtgag tttacttgta cagctcgtcc
      781 atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc
      841 tcgttggggt ctttgctcag ggcggactgg gtgctcaggt agtggttgtc gggcagcagc
      901 acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg
      961 tcctcgatgt tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc
     1021 atgatataga cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg
     1081 tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac
     1141 ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg
     1201 acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag
     1261 cggctgaagc actgcacgcc gtaggtgaag gtggtcacga gggtgggcca gggcacgggc
     1321 agcttgccgg tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg
     1381 ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg
     1441 ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca ttattcggag ttgtatttat
     1501 gttactaaaa caactggtca agttctacaa atacaaccgt tattggaatt tccccgatcg
     1561 ttcaaacatt tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgatgat
     1621 tatcatataa tttctgttga attacgttaa gcatgtaata tttaacatgt aatgcatgac
     1681 gttatttatg agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat
     1741 agaaaacaaa atatcgcgcg caaacttgga taaattatcg cgcgcggtgt catctatgag
     1801 gactagatcg gggtaccgag ctcgaattcg taatcatgtc atagctgttt cctgtgtgaa
     1861 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     1921 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1981 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2041 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2101 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2161 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2221 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2281 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2341 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2401 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2461 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2521 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2581 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2641 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2701 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2761 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2821 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2881 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2941 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3001 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3061 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3121 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3181 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3241 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3301 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3361 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3421 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3481 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3541 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3601 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3661 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3721 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3781 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3841 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     3961 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     4021 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     4081 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     4141 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     4201 ttaactatgc ggcatcagag ca
//
