LOCUS       sequence-124929-        5284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Promoterless AAV plasmid with MCS to insert promoter for expression 
            of iCre. Contains WPRE. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5284)
  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 5284)
  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: pEMS2159
FEATURES             Location/Qualifiers
     source          1..5284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          249..381
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     364..383
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     complement(1505..1525)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1923..1934)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2055..2176
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2066..2085)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2120..2139
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2345..2474
                     /label=AAV2 ITR
     primer_bind     complement(2493..2510)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2493..2509)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2502..2524)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2651..3106
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2738..2757)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2948..2969
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3132..3236
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3237..3305,3306..4097)
                     /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(3455..3474)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4268..4856
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4757..4776
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5144..5165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5180..5185,5186..5203,5204..5210)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5218..5234
                     /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     5223..5245
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5242..5258
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5242..5258
                     /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 gcgtggcgcg ccgaagttgg tcgtgaggca
      241 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      301 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      361 actttgcctt tctctccaca ggtgtccact cccaggcggc cgccaccatg tccaacctgc
      421 tgactgtgca ccaaaacctg cctgccctcc ctgtggatgc cacctctgat gaagtcagga
      481 agaacctgat ggacatgttc agggacaggc aggccttctc tgaacacacc tggaagatgc
      541 tcctgtctgt gtgcagatcc tgggctgcct ggtgcaagct gaacaacagg aaatggttcc
      601 ctgctgaacc tgaggatgtg agggactacc tcctgtacct gcaagccaga ggcctggctg
      661 tgaagaccat ccaacagcac ctgggccagc tcaacatgct gcacaggaga tctggcctgc
      721 ctcgcccttc tgactccaat gctgtgtccc tggtgatgag gagaatcaga aaggagaatg
      781 tggatgctgg ggagagagcc aagcaggccc tggcctttga acgcactgac tttgaccaag
      841 tcagatccct gatggagaac tctgacagat gccaggacat caggaacctg gccttcctgg
      901 gcattgccta caacaccctg ctgcgcattg ccgaaattgc cagaatcaga gtgaaggaca
      961 tctcccgcac cgatggtggg agaatgctga tccacattgg caggaccaag accctggtgt
     1021 ccacagctgg tgtggagaag gccctgtccc tgggggttac caagctggtg gagagatgga
     1081 tctctgtgtc tggtgtggct gatgacccca acaactacct gttctgccgg gtcagaaaga
     1141 atggtgtggc tgccccttct gccacctccc aactgtccac ccgggccctg gaagggatct
     1201 ttgaggccac ccaccgcctg atctatggtg ccaaggatga ctctgggcag agatacctgg
     1261 cctggtctgg ccactctgcc agagtgggtg ctgccaggga catggccagg gctggtgtgt
     1321 ccatccctga aatcatgcag gctggtggct ggaccaatgt gaacatagtg atgaactaca
     1381 tcagaaacct ggactctgag actggggcca tggtgaggct gctcgaggat ggggactgag
     1441 cggccgcgat cgctcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     1501 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     1561 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     1621 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     1681 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     1741 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     1801 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     1861 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     1921 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     1981 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     2041 gatcgcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     2101 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     2161 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     2221 gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtagtc gaaattcccg
     2281 ataaggatct tcctagagca tggctacgta gataagtagc atggcgggtt aatcattaac
     2341 tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     2401 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     2461 gagcgagcgc gcagccttaa ttaacctaat tcactggccg tcgttttaca acgtcgtgac
     2521 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     2581 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     2641 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2701 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2761 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2821 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2881 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2941 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3001 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3061 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     3121 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3181 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3241 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3301 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3361 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3421 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3481 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3541 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3601 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3661 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3721 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3781 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3841 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3901 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3961 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4021 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4081 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4141 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4201 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4261 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4321 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4381 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4441 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4501 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4561 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4621 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4681 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4741 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4801 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4861 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4921 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4981 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5041 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     5101 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     5161 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     5221 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc agatttaatt
     5281 aagg
//
