LOCUS       sequence-323553-        6998 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependently expresses the fusion protein EGFPL10a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6998)
  AUTHORS   Nectow AR, Moya MV, Ekstrand MI, Mousa A, McGuire KL, Sferrazza CE, 
            Field BC, Rabinowitz GS, Sawicka K, Liang Y, Friedman JM, Heintz N, 
            Schmidt EF
  TITLE     Rapid Molecular Profiling of Defined Cell Types Using Viral TRAP.
  JOURNAL   Cell Rep. 2017 Apr 18;19(3):655-667. doi: 
            10.1016/j.celrep.2017.03.048.
  PUBMED    28423326
REFERENCE   2  (bases 1 to 6998)
  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"; doi: 
            "10.1016/j.celrep.2017.03"; journalName: "Cell Rep"; date: 
            "2017-04-18- 18"; volume: "19"; issue: "3"; pages: "655-667"
COMMENT     Sequence Label: pAAV-FLEX-EGFPL10a
FEATURES             Location/Qualifiers
     source          1..6998
                     /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    complement(1440..1473)
                     /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)."
     protein_bind    complement(1524..1557)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(2256..2966,2967..2969,2970..2972))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(2256..2966,2967..2969,2970..2972))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2298..2319)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2647..2666
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2906..2927
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2969..2978
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2983..3016
                     /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)."
     protein_bind    3067..3100
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3125..3713
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3178..3198)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3596..3607)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3745..4221
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3944..3963)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4261..4401
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4261..4390
                     /label=AAV2 ITR
     rep_origin      4476..4931
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4563..4582)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4773..4794
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4948..4967)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5067..5089
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5127..5145)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5213..5317
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5318..5386,5387..6178)
                     /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(5536..5555)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6349..6937
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6838..6857
                     /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 gactccggaa
     1441 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
     1501 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
     1561 gcgccctaat acagacgctg gggcttgccc atggtgctct tgatgtacag agcccgcacg
     1621 ttttgccagt ttttcttaag caaggacacc aagaaattga cagccagatg aatgttgtag
     1681 actagctcat catcggtcat cttcacgtgg ccaacagcga cggccaaaca cagcaccttc
     1741 ttcatctgga acttgattgt cgatttcacc tcatccactt tggccaccat gttttcattg
     1801 tgtgtcagca gggaggggaa cttgccagcc ttgtttaggc ctgggcccag gatacgtggg
     1861 atctgcttaa tcagagactc agaggccaaa aaggcatcgt acttcttagc cagcttcttg
     1921 accaacttct tgtttttgtt aagcttcttg agcgcctcga tgtccatgtg ggggatatcc
     1981 acggccttgg cttcatcaca gtgctgctgg tcccccagaa cgcacaccga gaacttgggg
     2041 cgtggggtgg acttgagcct gacggtgccc gagaaacgtt tgtccttctg agggtcgtag
     2101 ttcttcaggc tgatctgcag ctccaccgtc tccagaaact tgcggcgctt gcgctggttc
     2161 ccgtgcagga cttcccgcac cgcctcgtac agggtgtcgc gtgagacttt gctgctgaat
     2221 tcgaagcttg agctcgagat ctgagtccgg ccggacttgt acagctcgtc catgccgaga
     2281 gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg
     2341 tctttgctca gggcggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg
     2401 tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg
     2461 ttgtggcgga tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag
     2521 acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg
     2581 aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg
     2641 gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct
     2701 tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag
     2761 cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg
     2821 gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg
     2881 gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc
     2941 ccggtgaaca gctcctcgcc cttgctcacc atggtggcta gcataacttc gtataaagta
     3001 tcctatacga agttatttgc cttaacccag aaattatcac tgttattctt tagaatggtg
     3061 caaagaataa cttcgtataa tgtatgctat acgaagttat gaattcgata tcaagcttat
     3121 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3181 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3241 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     3301 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     3361 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     3421 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     3481 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     3541 gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     3601 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     3661 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     3721 ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc
     3781 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     3841 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     3901 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     3961 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     4021 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     4081 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     4141 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     4201 actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc
     4261 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     4321 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     4381 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     4441 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     4501 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     4561 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4621 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4681 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4741 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4801 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     4861 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4921 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4981 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     5041 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     5101 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     5161 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     5221 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5281 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5341 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5401 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5461 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5521 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5581 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5641 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5701 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5761 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5821 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5881 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5941 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6001 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6061 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6121 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6181 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6241 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6301 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6361 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6421 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6481 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6541 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6601 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6661 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6721 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6781 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6841 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6901 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6961 acggttcctg gccttttgct ggccttttgc tcacatgt
//
