LOCUS       sequence-250828-        4216 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 2 attachment sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4216)
  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 4216)
  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)2
FEATURES             Location/Qualifiers
     source          1..4216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     66..88
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     80..97
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     81..97
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        179..523
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     467..487
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             complement(join(622..1335,1336..1338,1339..1341))
                     /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(667..688)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1016..1035
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1275..1296
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     terminator      1415..1668
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1697..1713)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1697..1713)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1710..1732)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1721..1737
                     /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(1745..1751,1752..1769,1770..1775))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1790..1811
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1928..1945)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2099..2687)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2179..2198)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2858..3649,3650..3718))
                     /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     3481..3500
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3719..3823)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3891..3909
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3947..3969)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4069..4088
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
       61 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagcttgcat gcctgcaggt
      121 cgactctaga ggatccccta attcatcaaa ttataactat atagacccta atttcatatg
      181 agacttttca acaaagggta atatccggaa acctcctcgg attccattgc ccagctatct
      241 gtcactttat tgtgaagata gtggaaaagg aaggtggctc ctacaaatgc catcattgcg
      301 ataaaggaaa ggccatcgtt gaagatgcct ctgccgacag tggtcccaaa gatggacccc
      361 cacccacgag gagcatcgtg gaaaaagaag acgttccaac cacgtcttca aagcaagtgg
      421 attgatgtga tatctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag
      481 acccttcctc tatataagga agttcatttc atttggagag gactaactgt gccaaattat
      541 cttccaaact cctaatccaa ttggacggcg cagaagggga gtagctcttc gccggaccgt
      601 cgacatactg ctcagctcgt cttacttgta cagctcgtcc atgccgagag tgatcccggc
      661 ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag
      721 ggcggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg
      781 ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt tgtggcggat
      841 cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga cgttgtggct
      901 gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc
      961 cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt
     1021 gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc
     1081 ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc actgcacgcc
     1141 gtaggtgaag gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat
     1201 gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa
     1261 cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag
     1321 ctcctcgccc ttgctcacca tagccaagag cagtgccttg cagaagttct cgggcgagaa
     1381 tctcgcgtag acacatacat gagcgaattt ccccgatcgt tcaaacattt ggcaataaag
     1441 tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt atcatataat ttctgttgaa
     1501 ttacgttaag catgtaatat ttaacatgta atgcatgacg ttatttatga gatgggtttt
     1561 tatgattaga gtcccgcaat tatacattta atacgcgata gaaaacaaaa tatcgcgcgc
     1621 aaacttggat aaattatcgc gcgcggtgtc atctatgagg actagatcgg ggtaccgagc
     1681 tcgaattcgt aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     1741 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     1801 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     1861 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     1921 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1981 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2041 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2101 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2161 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2221 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2281 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2341 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2401 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2461 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2521 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2581 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2641 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2701 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2761 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2821 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2881 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2941 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3001 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3061 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3121 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3181 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3241 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3301 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3361 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3421 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3481 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3541 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3601 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3661 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3721 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3781 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3841 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     3901 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     3961 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     4021 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     4081 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     4141 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     4201 tgcgggcctc ttcgct
//
