LOCUS       sequence-73209-e        4692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Promoterless AAV plasmid with MCS to insert promoter for expression 
            of iCre..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4692)
  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 4692)
  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: pEMS1980
FEATURES             Location/Qualifiers
     source          1..4692
                     /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"
     polyA_signal    1462..1583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1473..1492)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1527..1546
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   1753..1882
                     /label=AAV2 ITR
     primer_bind     complement(1901..1918)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1901..1917)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1910..1932)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2059..2514
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2146..2165)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2356..2377
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2540..2644
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2645..2713,2714..3505)
                     /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(2863..2882)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3676..4264
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4165..4184
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4552..4573
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4588..4593,4594..4611,4612..4618)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4626..4642
                     /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     4631..4653
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4650..4666
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4650..4666
                     /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 cgcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     1501 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     1561 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     1621 agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaggtcga
     1681 aattcccgat aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa
     1741 tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     1801 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     1861 cagtgagcga gcgagcgcgc agccttaatt aacctaattc actggccgtc gttttacaac
     1921 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     1981 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     2041 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2101 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2161 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2221 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     2281 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2341 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     2401 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     2461 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     2521 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2581 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2641 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2701 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2761 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     2821 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     2881 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2941 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3001 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3061 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3121 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3181 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3241 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3301 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3361 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3421 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3481 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3541 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3601 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3661 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3721 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3781 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3841 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3901 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3961 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4021 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4081 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4141 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4201 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4261 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4321 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     4381 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4441 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     4501 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     4561 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     4621 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccag
     4681 atttaattaa gg
//
