LOCUS       sequence-257203-        7043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tdTomato under control of eukaryotic Ef1a promoter after 
            Flp-dependent recombination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7043)
  AUTHORS   Sciolino NR, Hsiang M, Mazzone CM, Wilson LR, Plummer NW, Amin J, 
            Smith KG, McGee CA, Fry SA, Yang CX, Powell JM, Bruchas MR, Kravitz 
            AV, Cushman JD, Krashes MJ, Cui G, Jensen P
  TITLE     Natural locus coeruleus dynamics during feeding.
  JOURNAL   Sci Adv. 2022 Aug 19;8(33):eabn9134. doi: 10.1126/sciadv.abn9134. 
            Epub 2022 Aug 19.
  PUBMED    35984878
REFERENCE   2  (bases 1 to 7043)
  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 Adv."; 
            date: "2022-08-19"; volume: "8(33):eabn9134. doi"; pages: " 
            10.1126/sciadv.abn9134. Epub 2022 Aug 19"
COMMENT     Sequence Label: pAAV-Ef1a-fDIO-tdTomato
FEATURES             Location/Qualifiers
     source          1..7043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1440..1473
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(1584..3014)
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     regulatory      3011..3020
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3028..3061)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    3170..3758
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3223..3243)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3641..3652)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3790..4266
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3989..4008)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4306..4446
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4306..4435
                     /label=AAV2 ITR
     rep_origin      4521..4976
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4608..4627)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4818..4839
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4993..5012)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5112..5134
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5172..5190)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5258..5362
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5363..5431,5432..6223)
                     /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(5581..5600)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6394..6982
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6883..6902
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggag
     1441 aagttcctat tctctagaaa gtataggaac ttcgcagaat ggtagctgga ttgtagctgc
     1501 tattagcaat atgaaacctc ttagaagttc ctattcttca aaaggtatag gaacttcggc
     1561 gcgccggatc aagcttcgaa ttcttacttg tacagctcgt ccatgccgta caggaacagg
     1621 tggtggcggc cctcggagcg ctcgtactgt tccacgatgg tgtagtcctc gttgtgggag
     1681 gtgatgtcca gcttggtgtc cacgtagtag tagccgggca gttgcacggg cttcttggcc
     1741 atgtagatgg tcttgaactc caccaggtag tggccgccgt ccttcagctt cagggcctgg
     1801 tggatctcgc ccttcagcac gccgtcgcgg gggtacaggc gctcggtgga ggcctcccag
     1861 cccatggtct tcttctgcat tacggggccg tcggggggga agttggtgcc gcgcatcttc
     1921 accttgtaga tcagcgtgcc gtcctgcagg gaggagtcct gggtcacggt caccagaccg
     1981 ccgtcctcga agttcatcac gcgctcccac ttgaagccct cggggaagga cagcttcttg
     2041 taatcgggga tgtcggcggg gtgcttcacg tacgccttgg agccgtacat gaactggggg
     2101 gacaggatgt cccaggcgaa gggcaggggg ccgcccttgg tcaccttcag cttggcggtc
     2161 tgggtgccct cgtaggggcg gccctcgccc tcgccctcga tctcgaactc gtggccgttc
     2221 atggagccct ccatgcgcac cttgaagcgc atgaactctt tgatgacggc catgttgttg
     2281 tcctcggagg aggcggtgcc ggagctgccg ctgccggtgc tgccggtgcc atgccccagg
     2341 aacaggtggt ggcggccctc ggagcgctcg tactgttcca cgatggtgta gtcctcgttg
     2401 tgggaggtga tgtccagctt ggtgtccacg tagtagtagc cgggcagttg cacgggcttc
     2461 ttggccatgt agatggtctt gaactccacc aggtagtggc cgccgtcctt cagcttcagg
     2521 gcctggtgga tctcgccctt cagcacgccg tcgcgggggt acaggcgctc ggtggaggcc
     2581 tcccagccca tggtcttctt ctgcattacg gggccgtcgg gggggaagtt ggtgccgcgc
     2641 atcttcacct tgtagatcag cgtgccgtcc tgcagggagg agtcctgggt cacggtcacc
     2701 agaccgccgt cctcgaagtt catcacgcgc tcccacttga agccctcggg gaaggacagc
     2761 ttcttgtaat cggggatgtc ggcggggtgc ttcacgtacg ccttggagcc gtacatgaac
     2821 tggggggaca ggatgtccca ggcgaagggc agggggccgc ccttggtcac cttcagcttg
     2881 gcggtctggg tgccctcgta ggggcggccc tcgccctcgc cctcgatctc gaactcgtgg
     2941 ccgttcatgg agccctccat gcgcaccttg aagcgcatga actctttgat gacctcctcg
     3001 cccttgctca ccatggtggc ggctagcgaa gttcctatac tttctagaga ataggaactt
     3061 cttgccttaa cccagaaatt atcactgtta ttctttagaa tggtgcaaag agaagttcct
     3121 ataccttttg aagaatagga acttcgaatt cgatatcaag cttatcgata atcaacctct
     3181 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3241 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3301 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3361 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3421 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3481 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3541 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc
     3601 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3661 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3721 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga
     3781 gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     3841 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     3901 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     3961 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     4021 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     4081 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     4141 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     4201 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     4261 gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga
     4321 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     4381 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     4441 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4501 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4561 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     4621 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4681 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     4741 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4801 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     4861 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     4921 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     4981 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     5041 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     5101 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     5161 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     5221 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5281 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5341 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5401 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5461 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5521 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5581 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5641 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5701 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5761 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5821 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5881 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5941 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6001 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6061 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6121 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6181 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6241 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6301 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6361 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6421 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6481 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6541 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6601 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6661 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6721 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6781 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6841 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6901 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6961 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7021 ttgctggcct tttgctcaca tgt
//
