LOCUS       sequence-73234-e        5894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid with TNNT1-B (Ple301) promoter driving expression of 
            iCre..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5894)
  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 5894)
  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: pEMS2026
FEATURES             Location/Qualifiers
     source          1..5894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          1452..1584
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1567..1586
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     polyA_signal    2665..2786
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2676..2695)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2730..2749
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2955..3084
                     /label=AAV2 ITR
     primer_bind     complement(3103..3120)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3103..3119)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3112..3134)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3261..3716
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3348..3367)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3558..3579
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3742..3846
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3847..3915,3916..4707)
                     /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(4065..4084)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4878..5466
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5367..5386
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5754..5775
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5790..5795,5796..5813,5814..5820)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5828..5844
                     /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     5833..5855
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5852..5868
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5852..5868
                     /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 ggccggccga aaacttagag tgccattatc tggggtatca
      241 tgaactgggc ctccgagttt gggtatcaag attttggggg tctcatattc ttgaggtttc
      301 agattttagg aggtctcaag acctgggggt ggtcttagga tgtcccggcc catatctgga
      361 ctatcaagaa ctgggatttc agaatccaga gctgtgagga cttgctggcc tcagaaacac
      421 gggcatcata aatcggggtc tcagaatctg tgggtattgg gatttggggc taggcaccct
      481 gtggcctagg gtgtctcaaa gactttgagg tctaagaatg taggggtcag ggtgaggtct
      541 acggacagaa ggtgcccttt atgcccctgc cccctacgtg atgtatggga aggtgaaagg
      601 ggagacaggt ggggaggaaa ccagccagtg gggtggggcg agtgggagag gggatgcggg
      661 tgactaattc tcccagaagc cccccttcct caaatccctt cttaggaaat gtcctcggtt
      721 tcaccatcta tgacatcccc ccaaaatagc tcctgggggt ggggcagcta ttgtcttcag
      781 gccactgtcc cttctcaaat gcctcttcct aatcccaact ggatcaggtt cccatggact
      841 tgtcataaga caaaagagga cagctgtgct gagggggcag ggtctgcagc ctcctggctg
      901 tgccaggacc acacctacca aggtctgtcc tcatgcatgc tttaggacag ccggccccct
      961 ccctcagacc caagagtcca gacctgagcc ctcctccctc agacgcagga gtacaggccc
     1021 ccagcccgct cctccctcag actcaggagt ctagactccc agcctctccc ctccaaggac
     1081 ctactgtgtg ccgggcatgg ggacgtcaaa gcacagggag ctataaggga ggtagttgga
     1141 gcaccacggg agcctggggt gtggggtggg agcaggggcg gggctcctgg gccatgcaga
     1201 tggggttggc atggggttgg ggaggggccc acggaggctt ctggactctg agtgtggagc
     1261 caggggcagc agggctgtct ctgaactccc aggcttgtgt ttgagcaaag gaattctctc
     1321 cccctccctc tcccctcggg gcgggcccga gcctccagct ataaacttcc cggagcttct
     1381 gtgccctcag caaggctcag cctcaagatt cacagcaggc gcgccgaagt tggtcgtgag
     1441 gcactgggca ggtaagtatc aaggttacaa gacaggttta aggagaccaa tagaaactgg
     1501 gcttgtcgag acagagaaga ctcttgcgtt tctgataggc acctattggt cttactgaca
     1561 tccactttgc ctttctctcc acaggtgtcc actcccaggc ggccgccacc atgtccaacc
     1621 tgctgactgt gcaccaaaac ctgcctgccc tccctgtgga tgccacctct gatgaagtca
     1681 ggaagaacct gatggacatg ttcagggaca ggcaggcctt ctctgaacac acctggaaga
     1741 tgctcctgtc tgtgtgcaga tcctgggctg cctggtgcaa gctgaacaac aggaaatggt
     1801 tccctgctga acctgaggat gtgagggact acctcctgta cctgcaagcc agaggcctgg
     1861 ctgtgaagac catccaacag cacctgggcc agctcaacat gctgcacagg agatctggcc
     1921 tgcctcgccc ttctgactcc aatgctgtgt ccctggtgat gaggagaatc agaaaggaga
     1981 atgtggatgc tggggagaga gccaagcagg ccctggcctt tgaacgcact gactttgacc
     2041 aagtcagatc cctgatggag aactctgaca gatgccagga catcaggaac ctggccttcc
     2101 tgggcattgc ctacaacacc ctgctgcgca ttgccgaaat tgccagaatc agagtgaagg
     2161 acatctcccg caccgatggt gggagaatgc tgatccacat tggcaggacc aagaccctgg
     2221 tgtccacagc tggtgtggag aaggccctgt ccctgggggt taccaagctg gtggagagat
     2281 ggatctctgt gtctggtgtg gctgatgacc ccaacaacta cctgttctgc cgggtcagaa
     2341 agaatggtgt ggctgcccct tctgccacct cccaactgtc cacccgggcc ctggaaggga
     2401 tctttgaggc cacccaccgc ctgatctatg gtgccaagga tgactctggg cagagatacc
     2461 tggcctggtc tggccactct gccagagtgg gtgctgccag ggacatggcc agggctggtg
     2521 tgtccatccc tgaaatcatg caggctggtg gctggaccaa tgtgaacata gtgatgaact
     2581 acatcagaaa cctggactct gagactgggg ccatggtgag gctgctcgag gatggggact
     2641 gagcggccgc gatcgcagac atgataagat acattgatga gtttggacaa accacaacta
     2701 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     2761 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     2821 ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtagtc
     2881 gaaattcccg ataaggatct tcctagagca tggctacgta gataagtagc atggcgggtt
     2941 aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     3001 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     3061 ctcagtgagc gagcgagcgc gcagccttaa ttaacctaat tcactggccg tcgttttaca
     3121 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     3181 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     3241 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3301 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3361 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3421 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3481 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3541 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3601 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3661 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
     3721 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3781 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3841 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3901 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3961 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4021 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4081 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4141 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4201 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     4261 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     4321 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     4381 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     4441 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     4501 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     4561 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     4621 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     4681 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     4741 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4801 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4861 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4921 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4981 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5041 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5101 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5161 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5221 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5281 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5341 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5401 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5461 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5521 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     5581 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     5641 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     5701 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     5761 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     5821 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     5881 agatttaatt aagg
//
