LOCUS       sequence-322507-        6348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent EF1a-driven expression of mGreenLantern in animals 
            using AAV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6348)
  AUTHORS   Campbell BC, Nabel EM, Murdock MH, Lao-Peregrin C, Tsoulfas P, 
            Blackmore MG, Lee FS, Liston C, Morishita H, Petsko GA
  TITLE     mGreenLantern: a bright monomeric fluorescent protein with rapid 
            expression and cell filling properties for neuronal imaging.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Dec 1;117(48):30710-30721. doi: 
            10.1073/pnas.2000942117. Epub 2020 Nov 18.
  PUBMED    33208539
REFERENCE   2  (bases 1 to 6348)
  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: 
            "10.1073/pnas.2000942117"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2020-12-1- 1"; volume: "117"; issue: "48"; pages: 
            "30710-30721"
COMMENT     Sequence Label: pAAV-EF1a-DIO-mGreenLantern
FEATURES             Location/Qualifiers
     source          1..6348
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1210..1243)
                     /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(1294..1327)
                     /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(1355..2068,2069..2071,2072..2074))
                     /codon_start=1
                     /product="monomeric GFP derivative that displays 
                     exceptionally high brightness in cells (Campbell et al., 
                     2020)"
                     /label=mGreenLantern
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLGYGVACFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIVLKGIDFKEDGNILGHKLEYNFNSHKVYITADKQKNGIK
                     ANFKTRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLK
                     ERVTAAGITHDMDELYK"
     primer_bind     2008..2029
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2071..2080
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2103..2136
                     /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    2187..2220
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2245..2833
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2298..2318)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2716..2727)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2865..3341
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3064..3083)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3381..3521
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3381..3510
                     /label=AAV2 ITR
     rep_origin      3596..4051
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3683..3702)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3893..3914
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4068..4087)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4187..4209
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4247..4265)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4333..4437
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4438..4506,4507..5298)
                     /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(4656..4675)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5469..6057
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5958..5977
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6119..6248
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc
     1201 gactccggaa taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga
     1261 ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg
     1321 aagttatggc gcgccttatc tagatttagc caccttactt gtacagctcg tccatgtcat
     1381 gtgtaatccc ggcggcggtc accctctcct tcaggaccat gtgatcgcgc ttctcgttgg
     1441 ggtctttgct cagcttggac tgatggctca ggtagtggtt gtcgggcagc agcacggggc
     1501 cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgccg ccgtcctcaa
     1561 cgttgtggcg ggtcttgaag ttagccttga tgccgttctt ctgcttgtcg gccgtgatat
     1621 agaccttgtg gctgttgaag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct
     1681 tgaagtcgat gcccttcagc acgatgcggt tcaccagggt gtcgccctcg aacttcacct
     1741 cggcgcgggt cttgtaggta ccgtcgtcct tgaaagagat ggtgcgctcc tggacgtagc
     1801 cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcgggcga
     1861 agcaggccac gccgtagcct aaggtggtca cgagggtggg ccagggcacg ggcagcttgc
     1921 cggtggtgca gatgaacttc agggtcagct tgccgttggt ggcatcgccc tcgccctcgc
     1981 cgcggacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca
     2041 ccccggtgaa cagctcctcg cccttgctca ccatggtggc ggtggcgacc ggtagcgcta
     2101 gcataacttc gtataaagta tcctatacga agttatttgc cttaacccag aaattatcac
     2161 tgttattctt tagaatggtg caaagaataa cttcgtataa tgtatgctat acgaagttat
     2221 gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     2281 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     2341 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     2401 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     2461 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2521 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2581 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     2641 tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc
     2701 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     2761 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     2821 gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg
     2881 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg
     2941 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg
     3001 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg
     3061 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg
     3121 cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat
     3181 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg
     3241 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc
     3301 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca
     3361 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     3421 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     3481 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     3541 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     3601 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3661 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3721 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3781 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     3841 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3901 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     3961 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4021 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     4081 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4141 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4201 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     4261 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4321 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4381 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4441 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4501 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4561 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4621 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4681 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4741 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4801 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4861 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4921 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4981 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5041 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5101 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5161 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5221 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5281 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5341 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5401 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5461 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5521 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5581 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5641 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5701 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5761 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5821 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5881 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5941 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6001 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6061 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc
     6121 tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg
     6181 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     6241 gggttcctgc ggccgcacgc gtaagctttg caaagatgga taaagtttta aacagagagg
     6301 aatctttgca gctaatggac cttctaggtc ttgaaaggag tgggaatt
//
