LOCUS       sequence-250746-        5187 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 7 attachment sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5187)
  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 5187)
  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)7
FEATURES             Location/Qualifiers
     source          1..5187
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        68..1250
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          299..1241
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1198..1218
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             complement(join(1368..2081,2082..2084,2085..2087))
                     /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(1371..2081,2082..2084,2085..2087))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1413..1434)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1762..1781
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2021..2042
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2241..2260)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2306..2361
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2360..2379)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2722..2738)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2722..2738)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2735..2757)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2746..2762
                     /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(2770..2776,2777..2794,2795..2800))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2815..2836
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2895..3090
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2941..3076
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3096..3230
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3133..3152)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3187..3206
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3298..3315)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3469..4057)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3549..3568)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4228..5019,5020..5088))
                     /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     4851..4870
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5089..5187,1..6))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca ttgattattg actagatcat
       61 cgcgtgaggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa
      121 gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg
      181 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
      241 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt
      301 aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc
      361 ttgaattact tccacgcccc tggctgcagt acgtgattct tgatcccgag cttcgggttg
      421 gaagtgggtg ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag
      481 ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct
      541 gtctcgctgc tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc
      601 tttttttctg gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg
      661 tttttggggc cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc
      721 ggggcctgcg agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg
      781 ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc
      841 ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct
      901 caaaatggag gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa
      961 gggcctttcc gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca
     1021 ggcacctcga ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt
     1081 tttatgcgat ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc
     1141 acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca
     1201 agcctcagac agtggttcaa agtttttttc ttccatttca ggtgtcgtga ggaattctgc
     1261 agtcgacggt accgcgggcc cgggatccac cggtcgccac cagacgagaa acgttccgtc
     1321 cgtctgggtc agttgggcaa agttgatgac cgggtcgtcc gttcctttta cttgtacagc
     1381 tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg
     1441 cgcttctcgt tggggtcttt gctcagggcg gactgggtgc tcaggtagtg gttgtcgggc
     1501 agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg
     1561 ctgccgtcct cgatgttgtg gcggatcttg aagttcacct tgatgccgtt cttctgcttg
     1621 tcggccatga tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg
     1681 ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc
     1741 tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc
     1801 tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg
     1861 gggtagcggc tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc
     1921 acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg
     1981 ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc
     2041 aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccataag ctgaacctgc
     2101 gtaaaagtta aggttaggca tgtaaactta actctcacac aggtttataa caccagcggc
     2161 cgcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg
     2221 gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag
     2281 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     2341 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
     2401 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
     2461 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
     2521 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
     2581 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
     2641 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag
     2701 cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2761 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     2821 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2881 gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     2941 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3001 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3061 ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg
     3121 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3181 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta
     3241 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3301 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3361 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3421 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3481 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3541 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3601 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3661 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3721 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3781 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3841 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3901 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3961 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4021 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4081 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4141 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4201 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4261 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4321 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4381 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4441 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4501 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4561 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4621 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4681 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4741 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4801 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4861 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4921 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4981 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5041 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5101 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5161 tatttagaaa aataaacaaa taggggt
//
