LOCUS       sequence-438337-        6291 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Cre-dependent expression of the red fluorescent protein mCherry 
            under control of the EF1a promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6291)
  AUTHORS   Blockus H, Rolotti SV, Szoboszlay M, Peze-Heidsieck E, Ming T, 
            Schroeder A, Apostolo N, Vennekens KM, Katsamba PS, Bahna F, 
            Mannepalli S, Ahlsen G, Honig B, Shapiro L, de Wit J, Losonczy A, 
            Polleux F
  TITLE     Synaptogenic activity of the axon guidance molecule Robo2 underlies 
            hippocampal circuit function.
  JOURNAL   Cell Rep. 2021 Oct 19;37(3):109828. doi: 
            10.1016/j.celrep.2021.109828.
  PUBMED    34686348
REFERENCE   2  (bases 1 to 6291)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep.";
            date: "2021-10-19"; volume: "37(3):109828. doi"; pages: " 
            10.1016/j.celrep.2021.109828"
COMMENT     Sequence Label: pEF1-alpha:flex-mcherry
FEATURES             Location/Qualifiers
     source          1..6291
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     131..150
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    381..969
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             487..498
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     896..916
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    complement(994..1027)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(1078..1111)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     regulatory      1113..1122
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1119..1829
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1264..1282)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1532..1551
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    1830..1863
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1914..1947
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     promoter        complement(1978..3156)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1987..2925
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(2010..2030)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     repeat_region   3257..3386
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3448..4036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3528..3547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4207..4998,4999..5067))
                     /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     4830..4849
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5068..5172)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5240..5258
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5296..5318)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5418..5437
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5454..5909)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5591..5612)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5803..5822
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5984..6124
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5995..6124
                     /label=AAV2 ITR
     polyA_signal    join(6164..6291,1..349)
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
ORIGIN
        1 aaaaaaacaa aaattagctg agcctggtca tgcatgcctg gaatcccaac aactcgggag
       61 gctgaggcag gagaatcgct tgaacccagg aggcggagat tgcagtgagc caagattgtg
      121 ccactgcact ccagcttggt tcccaataga ccccgcaggc cctacaggtt gtcttcccaa
      181 cttgcccctt gctccatacc acccccctcc accccataat attatagaag gacacctagt
      241 cagacaaaat gatgcaactt aattttatta ggacaaggct ggtgggcact ggagtggcaa
      301 cttccagggc caggagaggc actggggagg ggtcacaggg atgccacccg tagatctctc
      361 gagcagcgct cggtatcgat gcggggaggc ggcccaaagg gagatccgac tcgtctgagg
      421 gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg aaggaaggtc
      481 cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc
      541 aacataggcg agcagccaag gaaaggacga tgatttcccc gacaacacca cggaattgtc
      601 agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga tgagttccgc
      661 cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg tggtggcaat
      721 gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac gttgcctgac
      781 aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg aggagaaaat
      841 gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt atccacatag
      901 cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt tgtaatccag
      961 aggttgatta tcgataagct tgatatcgaa ttcataactt cgtatagcat acattatacg
     1021 aagttattct ttgcaccatt ctaaagaata acagtgataa tttctgggtt aaggcaaata
     1081 acttcgtata ggatacttta tacgaagtta tcgccaccat ggtgagcaag ggcgaggagg
     1141 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
     1201 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     1261 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
     1321 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
     1381 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     1441 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
     1501 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
     1561 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
     1621 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
     1681 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
     1741 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca
     1801 ccggcggcat ggacgagctg tacaagtaaa taacttcgta taatgtatgc tatacgaagt
     1861 tattaagagg tttcatattg ctaatagcag ctacaatcca gctaccattc tgcataactt
     1921 cgtataaagt atcctatacg aagttattcc ggagtcgact ctagaggatc cggtacctca
     1981 cgacacctga aatggaagaa aaaaactttg aaccactgtc tgaggcttga gaatgaacca
     2041 agatccaaac tcaaaaaggg caaattccaa ggagaattac atcaagtgcc aagctggcct
     2101 aacttcagtc tccacccact cagtgtgggg aaactccatc gcataaaacc cctcccccca
     2161 acctaaagac gacgtactcc aaaagctcga gaactaatcg aggtgcctgg acggcgcccg
     2221 gtactccgtg gagtcacatg aagcgacggc tgaggacgga aaggcccttt tcctttgtgt
     2281 gggtgactca cccgcccgct ctcccgagcg ccgcgtcctc cattttgagc tccctgcagc
     2341 agggccggga agcggccatc tttccgctca cgcaactggt gccgaccggg ccagccttgc
     2401 cgcccagggc ggggcgatac acggcggcgc gaggccaggc accagagcag gccggccagc
     2461 ttgagactac ccccgtccga ttctcggtgg ccgcgctcgc aggccccgcc tcgccgaaca
     2521 tgtgcgctgg gacgcacggg ccccgtcgcc gcccgcggcc ccaaaaaccg aaataccagt
     2581 gtgcagatct tggcccgcat ttacaagact atcttgccag aaaaaaagcg tcgcagcagg
     2641 tcatcaaaaa ttttaaatgg ctagagactt atcgaaagca gcgagacagg cgcgaaggtg
     2701 ccaccagatt cgcacgcggc ggccccagcg cccaggccag gcctcaactc aagcacgagg
     2761 cgaaggggct ccttaagcgc aaggcctcga actctcccac ccacttccaa cccgaagctc
     2821 gggatcaaga atcacgtact gcagccaggt ggaagtaatt caaggcacgc aagggccata
     2881 acccgtaaag aggccaggcc cgcgggaacc acacacggca cttacctgtg ttctggcggc
     2941 aaacccgttg cgaaaaagaa cgttcacggc gactactgca cttatatacg gttctccccc
     3001 accctcggga aaaaggcgga gccagtacac gacatcactt tcccagttta ccccgcgcca
     3061 ccttctctag gcaccggttc aattgccgac ccctcccccc aacttctcgg ggactgtggg
     3121 cgatgtgcgc tctgcccact gacgggcacc ggagccaatt cccactcctt tcaagaccta
     3181 gaaggtccat tagctgcaaa gattcctctc tgtttaaaac tttatccatc tttgcaaagc
     3241 ttacgcgtgc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3301 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag
     3361 cgagcgagcg cgcagctgcc tgcaggacat gtgagcaaaa ggccagcaaa aggccaggaa
     3421 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3481 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3541 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3601 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3661 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3721 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3781 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3841 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3901 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3961 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4021 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4081 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4141 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4201 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4261 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4321 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4381 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4441 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4501 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4561 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4621 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4681 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4741 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4801 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4861 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4921 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4981 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5041 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5101 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5161 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     5221 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     5281 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     5341 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     5401 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc ataaaattgt
     5461 aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     5521 ccaatagacc gaaatcggca aaatccctta taaatcaaaa gaatagcccg agatagagtt
     5581 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     5641 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac ccaaatcaag
     5701 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     5761 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     5821 agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     5881 cgcgcttaat gcgccgctac agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa
     5941 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gcccctgcag gcagctgcgc
     6001 gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc
     6061 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt
     6121 tcctgcggcc gctcggtccg cacgtggtta cctacaaaat cagaaggaca gggaagggag
     6181 cagtggttca cgcctgtaat cccagcaatt tgggaggcca aggtgggtag atcacctgag
     6241 attaggagtt ggagaccagc ctggccaata tggtgaaacc ccgtctctac c
//
