LOCUS       sequence-73218-e        7005 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid with CAGGS promoter driving expression of iCre. Contains
            WPRE..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7005)
  AUTHORS   Simpson E, Korecki AJ, Fornes O, McGill TJ, Cueva-Vargas JL, 
            Agostinone J, Farkas RA, Hickmott JW, Lam SL, Mathelier A, Renner 
            LM, Stoddard J, Zhou M, Di Polo A, Neuringer M, Wasserman WW
  TITLE     New MiniPromoter Ple345 (NEFL) Drives Strong and Specific Expression
            in Retinal Ganglion Cells of Mouse and Primate Retina.
  JOURNAL   Hum Gene Ther. 2018 Jul 31. doi: 10.1089/hum.2018.118.
  PUBMED    30062914
REFERENCE   2  (bases 1 to 7005)
  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: "Hum Gene 
            Ther. 2018 Jul 31. doi: 10.1089/hum.2018.118."
COMMENT     Sequence Label: pEMS1989
FEATURES             Location/Qualifiers
     source          1..7005
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     enhancer        216..595
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        598..873
                     /label=chicken beta-actin promoter
     intron          874..1882
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1806..1828
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     intron          1970..2102
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2085..2104
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     complement(3226..3246)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3644..3655)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3776..3897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3787..3806)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3841..3860
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4066..4195
                     /label=AAV2 ITR
     primer_bind     complement(4214..4231)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4214..4230)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4223..4245)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4372..4827
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4459..4478)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4669..4690
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4853..4957
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4958..5026,5027..5818)
                     /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(5176..5195)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5989..6577
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6478..6497
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6865..6886
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6901..6906,6907..6924,6925..6931)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6939..6955
                     /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     6944..6966
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6963..6979
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6963..6979
                     /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 gcgttgacat tgattattga ctagttatta
      241 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      301 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      361 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      421 ctatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      481 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      541 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatgggtcg
      601 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
      661 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggcgc
      721 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
      781 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
      841 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgttgcc ttcgccccgt
      901 gccccgctcc gcgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
      961 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga
     1021 cggctcgttt cttttctgtg gctgcgtgaa agccttaaag ggctccggga gggccctttg
     1081 tgcggggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg
     1141 cggcccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc
     1201 cgcgtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg ggggctgcga
     1261 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc
     1321 ggcggtcggg ctgtaacccc cccctgcacc cccctccccg agttgctgag cacggcccgg
     1381 cttcgggtgc ggggctccgt gcggggcgtg gcgcggggct cgccgtgccg ggcggggggt
     1441 ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg
     1501 aggggcgcgg cggccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
     1561 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctggcggag
     1621 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcgggcgaag cggtgcggcg
     1681 ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc
     1741 tccatctcca gcctcggggc tgccgcaggg ggacggctgc cttcgggggg gacggggcag
     1801 ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca
     1861 tgccttcttc tttttcctac agctcctggg caacgtgctg gttgttgtgc tgtctcatca
     1921 ttttggcaaa gaattggcgc gccgaagttg gtcgtgaggc actgggcagg taagtatcaa
     1981 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
     2041 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     2101 aggtgtccac tcccaggcgg ccgccaccat gtccaacctg ctgactgtgc accaaaacct
     2161 gcctgccctc cctgtggatg ccacctctga tgaagtcagg aagaacctga tggacatgtt
     2221 cagggacagg caggccttct ctgaacacac ctggaagatg ctcctgtctg tgtgcagatc
     2281 ctgggctgcc tggtgcaagc tgaacaacag gaaatggttc cctgctgaac ctgaggatgt
     2341 gagggactac ctcctgtacc tgcaagccag aggcctggct gtgaagacca tccaacagca
     2401 cctgggccag ctcaacatgc tgcacaggag atctggcctg cctcgccctt ctgactccaa
     2461 tgctgtgtcc ctggtgatga ggagaatcag aaaggagaat gtggatgctg gggagagagc
     2521 caagcaggcc ctggcctttg aacgcactga ctttgaccaa gtcagatccc tgatggagaa
     2581 ctctgacaga tgccaggaca tcaggaacct ggccttcctg ggcattgcct acaacaccct
     2641 gctgcgcatt gccgaaattg ccagaatcag agtgaaggac atctcccgca ccgatggtgg
     2701 gagaatgctg atccacattg gcaggaccaa gaccctggtg tccacagctg gtgtggagaa
     2761 ggccctgtcc ctgggggtta ccaagctggt ggagagatgg atctctgtgt ctggtgtggc
     2821 tgatgacccc aacaactacc tgttctgccg ggtcagaaag aatggtgtgg ctgccccttc
     2881 tgccacctcc caactgtcca cccgggccct ggaagggatc tttgaggcca cccaccgcct
     2941 gatctatggt gccaaggatg actctgggca gagatacctg gcctggtctg gccactctgc
     3001 cagagtgggt gctgccaggg acatggccag ggctggtgtg tccatccctg aaatcatgca
     3061 ggctggtggc tggaccaatg tgaacatagt gatgaactac atcagaaacc tggactctga
     3121 gactggggcc atggtgaggc tgctcgagga tggggactga gcggccgcga tcgctcaacc
     3181 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3241 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3301 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3361 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3421 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3481 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3541 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     3601 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3661 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3721 ccctcagacg agtcggatct ccctttgggc cgcctccccg cgatcgcaga catgataaga
     3781 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3841 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3901 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     3961 agcaagtaaa acctctacaa atgtggtagt cgaaattccc gataaggatc ttcctagagc
     4021 atggctacgt agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga
     4081 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     4141 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagcctta
     4201 attaacctaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     4261 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     4321 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     4381 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4441 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4501 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4561 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4621 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4681 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4741 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4801 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     4861 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4921 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4981 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5041 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5101 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5161 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5221 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5281 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5341 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5401 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5461 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5521 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5581 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5641 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5701 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5761 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5821 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5881 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5941 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6001 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6061 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6121 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6181 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6241 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6301 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6361 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6421 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6481 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6541 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6601 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6661 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     6721 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     6781 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     6841 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     6901 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     6961 cacaggaaac agctatgacc atgattacgc cagatttaat taagg
//
