LOCUS       sequence-73240-e        6582 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid with FEV (Ple67) promoter driving expression of EmGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6582)
  AUTHORS   Korecki AJ, Hickmott JW, Lam SL, Dreolini L, Mathelier A, Baker O, 
            Kuehne C, Bonaguro RJ, Smith J, Tan CV, Zhou M, Goldowitz D, 
            Deussing JM, Stewart AF, Wasserman WW, Holt RA, Simpson EM
  TITLE     Twenty-Seven Tamoxifen-Inducible iCre-Driver Mouse Strains for Eye 
            and Brain; Including Seventeen Carrying a New Inducible-First 
            Constitutive-Ready Allele.
  JOURNAL   Genetics. 2019 Feb 14. pii: genetics.119.301984. doi: 
            10.1534/genetics.119.301984.
  PUBMED    30765420
REFERENCE   2  (bases 1 to 6582)
  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: "Genetics. 
            2019 Feb 14. pii: genetics.119.301984. doi: 
            10.1534/genetics.119.301984."
COMMENT     Sequence Label: pEMS2114
FEATURES             Location/Qualifiers
     source          1..6582
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          2452..2584
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2567..2586
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2605..2614
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2611..2613,2614..2616,2617..3330)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2656..2677)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2917..2936)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3264..3285
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3353..3474
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3364..3383)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3418..3437
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3643..3772
                     /label=AAV2 ITR
     primer_bind     complement(3791..3808)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3791..3807)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3800..3822)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3949..4404
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4036..4055)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4246..4267
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4430..4534
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4535..4603,4604..5395)
                     /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(4753..4772)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5566..6154
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6055..6074
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6442..6463
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6478..6483,6484..6501,6502..6508)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6516..6532
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6521..6543
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6540..6556
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6540..6556
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatcg gaattcccta ggccggccac gcgccggaga agatcaagac tcaggtgggt
      241 ggagatgact gccccccatg cacacatttc ctggtgctcc accccctcag caaatgactt
      301 ctgggtaaga ctgagggata aagcctatac tgttgatcct tgcccagctc acagctggga
      361 tatttgggta gacctcacta ggaccctctc ctttacacct tagcctccca ctgagagcat
      421 taagtgggcg aattttgatg cctcccagct ttcccagctc ttacctcctg cccctggcta
      481 ttttctgttc ttacctcctg ctccttgttt tctgtccata gtgggtgggg gaagcagaaa
      541 gaggaatgag aaggagaggg attgggatct tgaccccaac ttcaaggccc tcaccttctg
      601 tggagccctg agtttggtcc acagggaaca ttccaacctc tagtcattgc tcccaccaga
      661 ttccaggtcc acatcctgaa ggcttcatcc ctgtcccact aggatcctcc caggccctgg
      721 gatccctctt tttccctcca cactctgcct cccactcttc tccctggctc ctttgcaccc
      781 tccacgagca aagatgaaca ctccagaagt tgcagcaaat gagggtgaac atgggccctg
      841 tagaacccca gaagtaggag tttccagcca tcctcaatca ggcatcccag gaccaggcac
      901 ttagcaggag acctgcccat tcccaggcct cctccacccc tgggctcaag cagtgagaga
      961 ggactttgag aatgccagtt ttgacagtac ccgttccata ctgcaaggct gcagaaggtg
     1021 ggcttctggg ctggagcctc atcttcagcc agcatcctcc ttagcctgcc tctccagagc
     1081 cacgaagccc cataggtacc cagagagact ccaggccact tgggaaccac atcaacatct
     1141 ttgcatgagt tagaaatagg aatccttttc ttggagtgga ttccagcctt gtacagtgca
     1201 caaccctgtc ttaacccctc cacctctcca gcctcgttca gggatagcca gaatgggagt
     1261 agtggtgagg gtgggtggca gactgggagg acaggggatg gggaaggatg ggagcagaat
     1321 agttgtacag ggaagtagca gggggctcag gaacccatca gggaaagcca ggacagattc
     1381 tcaacatctt tagaggcctt cagcagtcca agccactact aatttatggg gactgaatga
     1441 atgagaaaag gaggggaggg agaaggatga gagggagaga gaaggctgga ggggagatga
     1501 gagtcggaga gaaaagccta ggaggtaaag agagacctag aagggggcgg ggaggacaga
     1561 agagggggcc acccatcata agttgccagg agagatggcc cagcaccagg cctgaggctt
     1621 gggggcccca gaaaggcagg aaacgggcca aagcaagaag gagacgggga aggggttcgg
     1681 gtctctaggg aagagcagag agtgggagaa gaggaggagg gagcggggat gcgggcagag
     1741 ataaagggag caacggctgc agggccgggc gcgcagcgac aggcgagagg gagggaagcg
     1801 gaaatttaaa agtgaagatg cagataacgc agcctggaga cgggaacccg ggtggggata
     1861 gagggaggga gggggaggag agtgagcggc gagtggagcg ggagccgact ggaagaaggg
     1921 ctctagggag ggggctgtgg ctgctggggt ccgaggtggg gccgggtaca ccagccccat
     1981 cactgtttgc agagagtcag ggaggcggaa aagacacgcg ctctaggctc ccatcagggc
     2041 acatggcccg gacccatccc ccgcgcgtct ccccggctgc ggggcgcggg gggctgccgg
     2101 gtgcgcttgg ctgtggcgcg gcgcgttgga gactttattg cgatgggacg ataagagggg
     2161 cgggggcggg gtcctggggg ccgaggcggc agcgctttaa ttaaaacgga aattgcggcc
     2221 ccgggccgcg cgggggccgg agggttccaa gcggcccctt agctggaagc gtttctccag
     2281 gacccccccg caacccccgc cacgcccggg ctgccccctc ccgccaggcc ctgccggacc
     2341 cggcgccgtc ttctcctcct tgtcacccgc ggtcgcttcg ggcggggatc ggtgccaccg
     2401 agcgcaaagc ctgcctcggc gcgccgaagt tggtcgtgag gcactgggca ggtaagtatc
     2461 aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga
     2521 ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc
     2581 acaggtgtcc actcccaggc ggccgccacc atggtgagca agggcgagga gctgttcacc
     2641 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     2701 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     2761 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcaccta cggcgtgcag
     2821 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     2881 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     2941 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     3001 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaag
     3061 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg tgaacttcaa gacccgccac
     3121 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     3181 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     3241 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     3301 actctcggca tggacgagct gtacaagtaa gcggccgcga tcgcagacat gataagatac
     3361 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     3421 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3481 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc
     3541 aagtaaaacc tctacaaatg tggtagtcga aattcccgat aaggatcttc ctagagcatg
     3601 gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg
     3661 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     3721 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agccttaatt
     3781 aacctaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     3841 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     3901 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     3961 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4021 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4081 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4141 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4201 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4261 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4321 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4381 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4441 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4501 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4561 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4621 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4681 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4741 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4801 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4861 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4921 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4981 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5041 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5101 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5161 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5221 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5281 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5341 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5401 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5461 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5521 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5581 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5641 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5701 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5761 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5821 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5881 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5941 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6001 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6061 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6121 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6181 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6241 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     6301 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     6361 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     6421 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     6481 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     6541 aggaaacagc tatgaccatg attacgccag atttaattaa gg
//
