LOCUS       sequence-244155-        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     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        477..821
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     765..785
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             complement(join(920..1633,1634..1636,1637..1639))
                     /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(965..986)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1314..1333
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1573..1594
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     terminator      1713..1966
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1995..2011)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1995..2011)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2008..2030)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2019..2035
                     /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(2043..2049,2050..2067,2068..2073))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2088..2109
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2226..2243)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2397..2985)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2477..2496)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3156..3947,3948..4016))
                     /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     3779..3798
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4017..4121)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4189..4207
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttgcatgc ctgcaggtcg
      421 actctagagg atcccctaat tcatcaaatt ataactatat agaccctaat ttcatatgag
      481 acttttcaac aaagggtaat atccggaaac ctcctcggat tccattgccc agctatctgt
      541 cactttattg tgaagatagt ggaaaaggaa ggtggctcct acaaatgcca tcattgcgat
      601 aaaggaaagg ccatcgttga agatgcctct gccgacagtg gtcccaaaga tggaccccca
      661 cccacgagga gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat
      721 tgatgtgata tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac
      781 ccttcctcta tataaggaag ttcatttcat ttggagagga ctaactgtgc caaattatct
      841 tccaaactcc taatccaatt ggacggcgca gaaggggagt agctcttcgc cggaccgtcg
      901 acatactgct cagctcgtct tacttgtaca gctcgtccat gccgagagtg atcccggcgg
      961 cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcaggg
     1021 cggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg
     1081 tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct
     1141 tgaagttcac cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt
     1201 tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct
     1261 tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt
     1321 agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg
     1381 acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt
     1441 aggtgaaggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga
     1501 acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact
     1561 tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct
     1621 cctcgccctt gctcaccata gccaagagca gtgccttgca gaagttctcg ggcgagaatc
     1681 tcgcgtagac acatacatga gcgaatttcc ccgatcgttc aaacatttgg caataaagtt
     1741 tcttaagatt gaatcctgtt gccggtcttg cgatgattat catataattt ctgttgaatt
     1801 acgttaagca tgtaatattt aacatgtaat gcatgacgtt atttatgaga tgggttttta
     1861 tgattagagt cccgcaatta tacatttaat acgcgataga aaacaaaata tcgcgcgcaa
     1921 acttggataa attatcgcgc gcggtgtcat ctatgaggac tagatcgggg taccgagctc
     1981 gaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     2041 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     2101 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     2161 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2221 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2281 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2341 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2401 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2461 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2521 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2581 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2641 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2701 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2761 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2821 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2881 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2941 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3001 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3061 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3121 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3181 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3241 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3301 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3361 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3421 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3481 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3541 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3601 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3661 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3721 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     3781 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     3841 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     3901 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     3961 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4021 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4081 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4141 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     4201 acgaggccct ttcgtc
//
