LOCUS       sequence-250839-        4227 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 7 attachment sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4227)
  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 4227)
  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)7
FEATURES             Location/Qualifiers
     source          1..4227
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     207..229
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     221..238
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     222..238
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        320..664
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     608..628
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             complement(join(770..1483,1484..1486,1487..1489))
                     /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(815..836)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1164..1183
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1423..1444
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     terminator      1567..1820
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1849..1865)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1849..1865)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1862..1884)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1873..1889
                     /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(1897..1903,1904..1921,1922..1927))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1942..1963
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2080..2097)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2251..2839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2331..2350)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3010..3801,3802..3870))
                     /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     3633..3652
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3871..3975)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4043..4061
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4099..4121)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     join(4221..4227,1..13)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
       61 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
      121 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
      181 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
      241 caagcttgca tgcctgcagg tcgactctag aggatcccct aattcatcaa attataacta
      301 tatagaccct aatttcatat gagacttttc aacaaagggt aatatccgga aacctcctcg
      361 gattccattg cccagctatc tgtcacttta ttgtgaagat agtggaaaag gaaggtggct
      421 cctacaaatg ccatcattgc gataaaggaa aggccatcgt tgaagatgcc tctgccgaca
      481 gtggtcccaa agatggaccc ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa
      541 ccacgtcttc aaagcaagtg gattgatgtg atatctccac tgacgtaagg gatgacgcac
      601 aatcccacta tccttcgcaa gacccttcct ctatataagg aagttcattt catttggaga
      661 ggactaactg tgccaaatta tcttccaaac tcctaatcca attagacgag aaacgttccg
      721 tccgtctggg tcagttgggc aaagttgatg accgggtcgt ccgttccttt tacttgtaca
      781 gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat
      841 cgcgcttctc gttggggtct ttgctcaggg cggactgggt gctcaggtag tggttgtcgg
      901 gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca
      961 cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct
     1021 tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga
     1081 tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc
     1141 cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc
     1201 gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt
     1261 cggggtagcg gctgaagcac tgcacgccgt aggtgaaggt ggtcacgagg gtgggccagg
     1321 gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat
     1381 cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga
     1441 ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccata agctgaacct
     1501 gcgtaaaagt taaggttagg catgtaaact taactctcac acaggtttat aacaccgaat
     1561 ttccccgatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc tgttgccggt
     1621 cttgcgatga ttatcatata atttctgttg aattacgtta agcatgtaat atttaacatg
     1681 taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca attatacatt
     1741 taatacgcga tagaaaacaa aatatcgcgc gcaaacttgg ataaattatc gcgcgcggtg
     1801 tcatctatga ggactagatc ggggtaccga gctcgaattc gtaatcatgt catagctgtt
     1861 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     1921 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     1981 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2041 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2101 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2161 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2221 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2281 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2341 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2401 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2461 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2521 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2581 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2641 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2701 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2761 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2821 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2881 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2941 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3001 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3061 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3121 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3181 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3241 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3301 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3361 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3421 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3481 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3541 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3601 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3661 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     3721 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     3781 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3841 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3901 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3961 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     4021 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     4081 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     4141 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     4201 tcggggctgg cttaactatg cggcatc
//
