LOCUS       sequence-73239-e        7175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid with FEV (Ple67) promoter driving expression of EmGFP. 
            Contains WPRE..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7175)
  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 7175)
  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: pEMS2113
FEATURES             Location/Qualifiers
     source          1..7175
                     /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"
     primer_bind     complement(3396..3416)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3814..3825)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3946..4067
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3957..3976)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4011..4030
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4236..4365
                     /label=AAV2 ITR
     primer_bind     complement(4384..4401)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4384..4400)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4393..4415)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4542..4997
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4629..4648)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4839..4860
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5023..5127
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5128..5196,5197..5988)
                     /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(5346..5365)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6159..6747
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6648..6667
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    7035..7056
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7071..7076,7077..7094,7095..7101)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7109..7125
                     /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     7114..7136
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7133..7149
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7133..7149
                     /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 tcgctcaacc tctggattac
     3361 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     3421 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     3481 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     3541 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     3601 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     3661 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     3721 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     3781 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     3841 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     3901 agtcggatct ccctttgggc cgcctccccg cgatcgcaga catgataaga tacattgatg
     3961 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     4021 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     4081 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa
     4141 acctctacaa atgtggtagt cgaaattccc gataaggatc ttcctagagc atggctacgt
     4201 agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc
     4261 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     4321 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagcctta attaacctaa
     4381 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     4441 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     4501 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
     4561 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4621 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4681 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4741 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4801 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4861 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4921 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4981 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5041 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5101 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5161 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5221 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5281 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5341 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5401 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5461 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5521 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5581 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5641 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5701 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5761 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5821 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5881 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5941 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6001 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6061 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6121 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6181 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6241 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6301 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6361 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6421 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6481 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6541 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6601 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6661 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6721 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6781 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6841 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6901 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     6961 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     7021 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     7081 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     7141 agctatgacc atgattacgc cagatttaat taagg
//
