LOCUS       sequence-250744-        5183 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 4 attachment sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5183)
  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 5183)
  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)4
FEATURES             Location/Qualifiers
     source          1..5183
                     /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(2237..2256)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2302..2357
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2356..2375)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2718..2734)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2718..2734)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2731..2753)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2742..2758
                     /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(2766..2772,2773..2790,2791..2796))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2811..2832
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2891..3086
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2937..3072
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3092..3226
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3129..3148)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3183..3202
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3294..3311)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3465..4053)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3545..3564)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4224..5015,5016..5084))
                     /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     4847..4866
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5085..5183,1..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 cttccaaaga gcgcccaacg
     1321 cgacctgaaa tttgaataag actgctgctt gtgtaaaggc gatgatttta 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 tcaccataaa cgaattacaa
     2101 attatttgac cgctaattca aatcaagggt tgaaaacttt gtaatttttg agcggccgca
     2161 ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc
     2221 acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     2281 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     2341 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     2401 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     2461 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     2521 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     2581 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     2641 gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac ctgcagccca
     2701 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2761 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2821 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2881 cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     2941 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3001 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3061 tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta
     3121 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3181 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga
     3241 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3301 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3361 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3421 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3481 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3541 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3601 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3661 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3721 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3781 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3841 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3901 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3961 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4021 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4081 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4141 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4201 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4261 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4321 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4381 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4441 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4501 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4561 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4621 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4681 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4741 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4801 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4861 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4921 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4981 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5041 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5101 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5161 tagaaaaata aacaaatagg ggt
//
