LOCUS       sequence-250834-        4215 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 5 attachment sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4215)
  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 4215)
  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)5
FEATURES             Location/Qualifiers
     source          1..4215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     144..166
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     158..175
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     159..175
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        257..601
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     545..565
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             complement(join(702..1415,1416..1418,1419..1421))
                     /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(747..768)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1096..1115
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1355..1376
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     terminator      1492..1745
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1774..1790)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1774..1790)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1787..1809)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1798..1814
                     /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(1822..1828,1829..1846,1847..1852))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1867..1888
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2005..2022)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2176..2764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2256..2275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2935..3726,3727..3795))
                     /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     3558..3577
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3796..3900)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3968..3986
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4024..4046)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4146..4165
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
       61 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
      121 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa
      181 gcttgcatgc ctgcaggtcg actctagagg atcccctaat tcatcaaatt ataactatat
      241 agaccctaat ttcatatgag acttttcaac aaagggtaat atccggaaac ctcctcggat
      301 tccattgccc agctatctgt cactttattg tgaagatagt ggaaaaggaa ggtggctcct
      361 acaaatgcca tcattgcgat aaaggaaagg ccatcgttga agatgcctct gccgacagtg
      421 gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca
      481 cgtcttcaaa gcaagtggat tgatgtgata tctccactga cgtaagggat gacgcacaat
      541 cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga
      601 ctaactgtgc caaattatct tccaaactcc taatccaatt gagcgccgga tcagggagtg
      661 gacggcctgg gagcgctaca cgctgtggct gcggtcggtg cttacttgta cagctcgtcc
      721 atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc
      781 tcgttggggt ctttgctcag ggcggactgg gtgctcaggt agtggttgtc gggcagcagc
      841 acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg
      901 tcctcgatgt tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc
      961 atgatataga cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg
     1021 tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac
     1081 ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg
     1141 acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag
     1201 cggctgaagc actgcacgcc gtaggtgaag gtggtcacga gggtgggcca gggcacgggc
     1261 agcttgccgg tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg
     1321 ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg
     1381 ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca tcagatcagg gtgcgcaagt
     1441 tgtcaacgct cccaggagag ttatcgactt gcgtattagg ggaatttccc cgatcgttca
     1501 aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc gatgattatc
     1561 atataatttc tgttgaatta cgttaagcat gtaatattta acatgtaatg catgacgtta
     1621 tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata cgcgatagaa
     1681 aacaaaatat cgcgcgcaaa cttggataaa ttatcgcgcg cggtgtcatc tatgaggact
     1741 agatcggggt accgagctcg aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
     1801 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     1861 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     1921 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1981 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2041 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2101 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2161 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2221 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2281 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2341 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2401 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2461 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2521 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2581 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2641 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2701 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2761 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2821 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2881 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2941 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3001 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3061 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3121 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3181 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3241 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3301 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3361 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3421 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3481 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3541 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3601 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3661 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3721 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3781 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3841 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3901 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     3961 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     4021 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     4081 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     4141 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     4201 cagatgcgta aggag
//
