LOCUS       sequence-250745-        5175 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 5 attachment sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5175)
  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 5175)
  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)5
FEATURES             Location/Qualifiers
     source          1..5175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        55..1237
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          286..1228
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1185..1205
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             complement(join(1350..2063,2064..2066,2067..2069))
                     /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(1353..2063,2064..2066,2067..2069))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1395..1416)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1744..1763
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2003..2024
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2216..2235)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2281..2336
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2335..2354)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2697..2713)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2697..2713)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2710..2732)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2721..2737
                     /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(2745..2751,2752..2769,2770..2775))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2790..2811
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2870..3065
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2916..3051
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3071..3205
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3108..3127)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3162..3181
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3273..3290)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3444..4032)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3524..3543)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4203..4994,4995..5063))
                     /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     4826..4845
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5064..5168)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccccgaaaag tgccacctgg gtcgacattg attattgact agatcatcgc gtgaggctcc
       61 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
      121 gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
      181 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
      241 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt
      301 ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc
      361 acgcccctgg ctgcagtacg tgattcttga tcccgagctt cgggttggaa gtgggtggga
      421 gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc
      481 tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt
      541 cgataagtct ctagccattt aaaatttttg atgacctgct gcgacgcttt ttttctggca
      601 agatagtctt gtaaatgcgg gccaagatct gcacactggt atttcggttt ttggggccgc
      661 gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc
      721 gcggccaccg agaatcggac gggggtagtc tcaagctggc cggcctgctc tggtgcctgg
      781 cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt cggcaccagt
      841 tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca gggagctcaa aatggaggac
      901 gcggcgctcg ggagagcggg cgggtgagtc acccacacaa aggaaaaggg cctttccgtc
      961 ctcagccgtc gcttcatgtg actccacgga gtaccgggcg ccgtccaggc acctcgatta
     1021 gttctcgagc ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga
     1081 gtttccccac actgagtggg tggagactga agttaggcca gcttggcact tgatgtaatt
     1141 ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc ctcagacagt
     1201 ggttcaaagt ttttttcttc catttcaggt gtcgtgagga attctgcagt cgacggtacc
     1261 gcgggcccgg gatccaccgg tcgccaccga gcgccggatc agggagtgga cggcctggga
     1321 gcgctacacg ctgtggctgc ggtcggtgct tacttgtaca gctcgtccat gccgagagtg
     1381 atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct
     1441 ttgctcaggg cggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg
     1501 ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg
     1561 tggcggatct tgaagttcac cttgatgccg ttcttctgct tgtcggccat gatatagacg
     1621 ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
     1681 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
     1741 cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg
     1801 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac
     1861 tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
     1921 gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac
     1981 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
     2041 gtgaacagct cctcgccctt gctcaccatc agatcagggt gcgcaagttg tcaacgctcc
     2101 caggagagtt atcgacttgc gtattaggga gcggccgcac tcctcaggtg caggctgcct
     2161 atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt
     2221 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
     2281 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     2341 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
     2401 tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc
     2461 agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag
     2521 gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt
     2581 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
     2641 tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca
     2701 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     2761 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     2821 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt
     2881 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     2941 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3001 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3061 gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3121 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3181 caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga
     3241 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3301 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3361 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3421 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3481 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3541 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3601 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3661 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3721 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3781 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3841 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3901 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3961 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4021 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4081 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4141 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4201 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4261 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4321 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4381 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4441 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4501 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4561 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4621 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4681 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4741 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4801 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4861 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4921 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4981 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5041 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5101 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5161 gttccgcgca cattt
//
