LOCUS       sequence-250833-        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 4 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)4
FEATURES             Location/Qualifiers
     source          1..4222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     178..200
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     192..209
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     193..209
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        291..635
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     579..599
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             complement(join(741..1454,1455..1457,1458..1460))
                     /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(786..807)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1135..1154
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1394..1415
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     terminator      1533..1786
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1815..1831)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1815..1831)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1828..1850)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1839..1855
                     /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(1863..1869,1870..1887,1888..1893))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1908..1929
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2046..2063)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2217..2805)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2297..2316)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2976..3767,3768..3836))
                     /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     3599..3618
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3837..3941)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4009..4027
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4065..4087)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4187..4206
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
       61 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
      121 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
      181 agtcacgacg ttgtaaaacg acggccagtg ccaagcttgc atgcctgcag gtcgactcta
      241 gaggatcccc taattcatca aattataact atatagaccc taatttcata tgagactttt
      301 caacaaaggg taatatccgg aaacctcctc ggattccatt gcccagctat ctgtcacttt
      361 attgtgaaga tagtggaaaa ggaaggtggc tcctacaaat gccatcattg cgataaagga
      421 aaggccatcg ttgaagatgc ctctgccgac agtggtccca aagatggacc cccacccacg
      481 aggagcatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt ggattgatgt
      541 gatatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca agacccttcc
      601 tctatataag gaagttcatt tcatttggag aggactaact gtgccaaatt atcttccaaa
      661 ctcctaatcc aattttccaa agagcgccca acgcgacctg aaatttgaat aagactgctg
      721 cttgtgtaaa ggcgatgatt ttacttgtac agctcgtcca tgccgagagt gatcccggcg
      781 gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagg
      841 gcggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg
      901 gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc
      961 ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac gttgtggctg
     1021 ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc
     1081 ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg
     1141 tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg
     1201 gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg
     1261 taggtgaagg tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg
     1321 aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac
     1381 ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc
     1441 tcctcgccct tgctcaccat aaacgaatta caaattattt gaccgctaat tcaaatcaag
     1501 ggttgaaaac tttgtaattt ttgaatttcc ccgatcgttc aaacatttgg caataaagtt
     1561 tcttaagatt gaatcctgtt gccggtcttg cgatgattat catataattt ctgttgaatt
     1621 acgttaagca tgtaatattt aacatgtaat gcatgacgtt atttatgaga tgggttttta
     1681 tgattagagt cccgcaatta tacatttaat acgcgataga aaacaaaata tcgcgcgcaa
     1741 acttggataa attatcgcgc gcggtgtcat ctatgaggac tagatcgggg taccgagctc
     1801 gaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     1861 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     1921 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     1981 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2041 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2101 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2161 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2221 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2281 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2341 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2401 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2461 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2521 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2581 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2641 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2701 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2761 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2821 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     2881 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     2941 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3001 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3061 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3121 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3181 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3241 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3301 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3361 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3421 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3481 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3541 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     3601 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     3661 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     3721 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     3781 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     3841 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     3901 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     3961 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     4021 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     4081 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     4141 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     4201 attgtactga gagtgcacca ta
//
