LOCUS       sequence-250750-        5181 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid has an EF1 alpha 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 5181)
  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 5181)
  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-pEF-GFP(rc)9
FEATURES             Location/Qualifiers
     source          1..5181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        44..1226
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          275..1217
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1174..1194
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             complement(join(1343..2056,2057..2059,2060..2062))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1346..2056,2057..2059,2060..2062))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1388..1409)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1737..1756
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1996..2017
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2211..2230)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2276..2331
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2330..2349)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2692..2708)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2692..2708)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2705..2727)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2716..2732
                     /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(2740..2746,2747..2764,2765..2770))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2785..2806
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2865..3060
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2911..3046
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3066..3200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3103..3122)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3157..3176
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3268..3285)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3439..4027)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3519..3538)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4198..4989,4990..5058))
                     /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     4821..4840
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5059..5163)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gccacctggg tcgacattga ttattgacta gatcatcgcg tgaggctccg gtgcccgtca
       61 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
      121 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
      181 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
      241 tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg
      301 gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca cgcccctggc
      361 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc
      421 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
      481 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
      541 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
      601 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
      661 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
      721 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
      781 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
      841 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg
      901 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg
      961 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct
     1021 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
     1081 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     1141 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     1201 tttttcttcc atttcaggtg tcgtgaggaa ttctgcagtc gacggtaccg cgggcccggg
     1261 atccaccggt cgccaccttt atattgcgaa aaataattgg cgaacgaggt aactggatac
     1321 ctcatccgcc aattaaaatt tgttacttgt acagctcgtc catgccgaga gtgatcccgg
     1381 cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca
     1441 gggcggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg
     1501 gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga
     1561 tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag acgttgtggc
     1621 tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc
     1681 ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct
     1741 tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg
     1801 cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag cactgcacgc
     1861 cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga
     1921 tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga
     1981 acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca
     2041 gctcctcgcc cttgctcacc attaattgga agttcggaat aactatgcag atacctgata
     2101 cacacttcca acaaaaacaa ccacagcggc cgcactcctc aggtgcaggc tgcctatcag
     2161 aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc
     2221 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
     2281 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     2341 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     2401 acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat
     2461 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
     2521 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
     2581 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
     2641 cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat ggtcatagct
     2701 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     2761 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     2821 actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca
     2881 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     2941 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3001 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3061 aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3121 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3181 tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3241 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3301 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3361 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3421 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3481 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3541 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3601 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3661 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3721 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3781 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3841 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3901 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3961 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4021 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4081 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4141 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4201 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4261 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4321 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4381 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4441 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4501 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4561 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4621 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4681 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4741 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4801 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4861 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4921 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4981 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5041 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5101 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5161 gcgcacattt ccccgaaaag t
//
