LOCUS       sequence-368151-        5609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Electra1 in bacterial cells and Electra1 and mScarlet in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5609)
  AUTHORS   Papadaki S, Wang X, Wang Y, Zhang H, Jia S, Liu S, Yang M, Zhang D, 
            Jia JM, Koster RW, Namikawa K, Piatkevich KD
  TITLE     Dual-expression system for blue fluorescent protein optimization.
  JOURNAL   Sci Rep. 2022 Jun 17;12(1):10190. doi: 10.1038/s41598-022-13214-0.
  PUBMED    35715437
REFERENCE   2  (bases 1 to 5609)
  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: "Sci Rep."; 
            date: "2022-06-17"; volume: "12(1):10190. doi"; pages: " 
            10.1038/s41598-022-13214-0"
COMMENT     Sequence Label: pHybrid-Electra1
FEATURES             Location/Qualifiers
     source          1..5609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(54..73)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     264..285
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(489..508)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     608..630
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(668..686)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        754..858
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(859..927,928..1719)
                     /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     complement(1077..1096)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1890..2478
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2379..2398
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2689..3068
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3069..3272
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3223..3243
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3269..3293
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        3305..3423
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     CDS             3464..3481
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      3512..3521
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      4252..4338
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     polyA_signal    4362..4483
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4373..4392)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4427..4446
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4524..4734
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4546..4742
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4593..4728
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      4743..4752
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4749..5444
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(4882..4900)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    5449..5496
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(5576..5609,1..422)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
       61 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
      121 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
      181 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
      241 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
      301 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
      361 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
      421 ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
      481 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
      541 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
      601 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
      661 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
      721 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
      781 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
      841 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
      901 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
      961 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1021 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1081 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     1141 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1201 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1261 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     1321 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     1381 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     1441 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     1501 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     1561 gccggtgagc gtggctctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     1621 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     1681 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     1741 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     1801 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1861 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1921 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1981 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2041 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2101 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2161 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2221 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2281 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2341 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2401 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2461 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2521 tggccttttg ctcacatggc tcgacagatc ttcaatattg gccattagcc atattattca
     2581 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
     2641 ataatatgta catttatatt ggctcatgtc caatatgacc gccatgttgg cattgattat
     2701 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     2761 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     2821 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     2881 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     2941 tgccaagtcc gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     3001 agtacatgac cttacgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     3061 ttaccatggt gatgcggttt tggcagtaca ccaatgggcg tggatagcgg tttgactcac
     3121 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     3181 aacgggactt tccaaaatgt cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc
     3241 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcactagaa
     3301 gcttcaccac aattcagcaa attgtgaaca tcatcacgtt catctttccc tggttgccaa
     3361 tggcccattt tcctgtcagt aacgagaagg tcgcgaattc aggcgctttt tagactggtc
     3421 gtaacccgtt tttggctaac aggaggaatt aaccatgggg agccaccatc accatcacca
     3481 tgctagcctc gagaattcac gcgtggtacc agccaccatg gtgtctaagg gcgaagagct
     3541 gatcgaggaa aatatgcgta tgaaggtggt catggaaggt tcggtcaacg gccaccaatt
     3601 caaatgcaca ggtgaaggag aaggcagacc gtacgaggga gtgcaaacca tgaggatcaa
     3661 agtcatcgag ggaggacccc tgccatttgc ctttgacatt cttgccacgt cgttcctgtt
     3721 cggcagcaag acctttatca agtacccggc cgacatccct gatttcttca aacagtcctt
     3781 tcctgagggc tttacttggg aaagagttac gagatacgaa gatggtggag tcgtcaccgt
     3841 cacgcaggac accagccttg aggatggggg gctcgtctac aacgtcaagg tcagaggggt
     3901 aaactttccc tccaatggtc ccgtgatgca gaagaagacc gagggttggg agcctttcac
     3961 agagatgatg tatccagcaa atggtggtct gagaggatat accgacatcg cactgaaagt
     4021 tgatggtgat ggccatctgc acgccaacat cgtgacaact tacaggtcaa aaaagaccgt
     4081 cggggacatc aagatgcccg gtgtccatgc cgttgattac cgcctggaaa gggtcgaaga
     4141 gagtgacaat gaaacctacg tagtgctaag agaagtggca gttgccaaat acagcaacct
     4201 tggtggtggc atggacgagc tgtacaagta atctagagtc gacccaggca tcaaataaaa
     4261 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     4321 ctcctgagta ggacaaatgc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     4381 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     4441 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgggt
     4501 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4561 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4621 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4681 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4741 aagccaccat ggtgagcaag ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc
     4801 acatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     4861 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct
     4921 cctgggacat cctgtcccct cagttcatgt acggctccag ggccttcatc aagcaccccg
     4981 ccgacatccc cgactactat aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga
     5041 tgaacttcga ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca
     5101 ccctgatcta caaggtgaag ctccgcggca ccaacttccc tcctgacggc cccgtaatgc
     5161 agaagaagac aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc
     5221 tgaagggcga cattaagatg gccctgcgcc tgaaggacgg cggccgctac ctggcggact
     5281 tcaagaccac ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc
     5341 gcaagttgga catcacctcc cacaacgagg actacaccgt ggtggaacag tacgaacgct
     5401 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtgaccg gcaataaaaa
     5461 gacagaataa aacgcacggt gttgggtcgt ttgttcggat ccgggctggc gtaatagcga
     5521 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg
     5581 ccctgtagcg gcgcattaag cgcggcggg
//
