LOCUS       sequence-250840-        4221 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 9 attachment sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4221)
  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 4221)
  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)9
FEATURES             Location/Qualifiers
     source          1..4221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..36
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     28..45
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     29..45
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        127..471
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     415..435
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             complement(join(576..1289,1290..1292,1293..1295))
                     /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(621..642)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     970..989
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1229..1250
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     terminator      1368..1621
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1650..1666)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1650..1666)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1663..1685)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1674..1690
                     /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(1698..1704,1705..1722,1723..1728))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1743..1764
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1881..1898)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2052..2640)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2132..2151)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2811..3602,3603..3671))
                     /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     3434..3453
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3672..3776)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3844..3862
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3900..3922)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4022..4041
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttgcatgc
       61 ctgcaggtcg actctagagg atcccctaat tcatcaaatt ataactatat agaccctaat
      121 ttcatatgag acttttcaac aaagggtaat atccggaaac ctcctcggat tccattgccc
      181 agctatctgt cactttattg tgaagatagt ggaaaaggaa ggtggctcct acaaatgcca
      241 tcattgcgat aaaggaaagg ccatcgttga agatgcctct gccgacagtg gtcccaaaga
      301 tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa
      361 gcaagtggat tgatgtgata tctccactga cgtaagggat gacgcacaat cccactatcc
      421 ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga ctaactgtgc
      481 caaattatct tccaaactcc taatccaatt tttatattgc gaaaaataat tggcgaacga
      541 ggtaactgga tacctcatcc gccaattaaa atttgttact tgtacagctc gtccatgccg
      601 agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg
      661 gggtctttgc tcagggcgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg
      721 ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg
      781 atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata
      841 tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc
      901 ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc
      961 tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag
     1021 ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg
     1081 aagcactgca cgccgtaggt gaaggtggtc acgagggtgg gccagggcac gggcagcttg
     1141 ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg
     1201 ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc
     1261 accccggtga acagctcctc gcccttgctc accattaatt ggaagttcgg aataactatg
     1321 cagatacctg atacacactt ccaacaaaaa caaccacgaa tttccccgat cgttcaaaca
     1381 tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat
     1441 aatttctgtt gaattacgtt aagcatgtaa tatttaacat gtaatgcatg acgttattta
     1501 tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca
     1561 aaatatcgcg cgcaaacttg gataaattat cgcgcgcggt gtcatctatg aggactagat
     1621 cggggtaccg agctcgaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat
     1681 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     1741 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     1801 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     1861 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     1921 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     1981 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2041 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2101 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2161 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2221 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2281 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2341 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2401 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2461 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     2521 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2581 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2641 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2701 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     2761 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     2821 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     2881 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     2941 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3001 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3061 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3121 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3181 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3241 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3301 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3361 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     3421 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     3481 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     3541 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     3601 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     3661 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     3721 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     3781 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     3841 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     3901 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     3961 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     4021 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     4081 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
     4141 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     4201 gcaaggcgat taagttgggt a
//
