LOCUS       sequence-478577-        4714 bp    DNA     circular SYN 21-AUG-2025
DEFINITION  To express D2GFP from a Rat SARE-Arc minimal promoter. The Arc gene 
            contains the Rat Arc native SV40 3'UTR containing two introns.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4714)
  TITLE     Ploski Lab Plasmids 2025
REFERENCE   2  (bases 1 to 4714)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 21, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pAAV-Rat SARE-Arc Min Promoter-D2GFP-SV40 3'UTR
FEATURES             Location/Qualifiers
     source          1..4714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   91..220
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(282..870)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(362..381)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1041..1832,1833..1901))
                     /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     1664..1683
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1902..2006)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2074..2092
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2130..2152)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2252..2271
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2288..2743)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2425..2446)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2637..2656
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2818..2958
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2829..2958
                     /label=AAV2 ITR
     CDS             join(3609..3611,3612..3614,3615..4325,4326..4331,
                     4332..4454)
                     /codon_start=1
                     /product="EGFP destabilized by fusion to residues 422-461 
                     of mouse ornithine decarboxylase, giving an in vivo 
                     half-life of ~2 hours"
                     /label=d2EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     primer_bind     complement(3654..3675)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3915..3934)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4262..4283
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4549..4670
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4586..4605)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4640..4659
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tctcaattga cgcgtcccgg ggctacctta agagagcgcg tatttaaatc gctaccttag
       61 gaccgttata gttatcgact gaattgccgc aggaacccct agtgatggag ttggccactc
      121 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
      181 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg acatgtgagc aaaaggccag
      241 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      301 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      361 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      421 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      481 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      541 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      601 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      661 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      721 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      781 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      841 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      901 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      961 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     1021 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     1081 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1141 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     1201 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1261 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1321 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1381 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1441 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1501 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1561 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1621 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1681 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1741 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1801 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1861 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1921 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1981 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     2041 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     2101 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     2161 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     2221 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     2281 caccataaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     2341 agctcatttt ttaaccaata gaccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     2401 cccgagatag agttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     2461 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     2521 tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     2581 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     2641 aagaaagcga aaggagcggg cgctaaggcg ctggcaagtg tagcggtcac gctgcgcgta
     2701 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtactatgg ttgctttgac
     2761 gtatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgcccct
     2821 gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg
     2881 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
     2941 ccatcactag gggttcctgc ggcccgcgcc attaccctgt tatccctaat ttaaatctcg
     3001 atgctacctt aagagaggat atcctctgcg cgtagagcct tcctgcgtgg ggaagctcct
     3061 tgctgcgtca tggctcagct attctcagtc tctctccttt tatggtgccg gaagcaagcg
     3121 gcttctctgg tctctaagat ccctccgggt gggaggcgcg cagcagagca cattagtcac
     3181 tccgggctgt gaaggggcgg gtccttgagg gcacccgcgg aaggggagcg agtaggcgcg
     3241 ggagaggcgg ggcctgcggc aggagagggc gcgggcgggc tctggcgcgg agcctgggcg
     3301 cggccaatgg gagccagggc tccgcgagct gccgcccacg ggccccgcgc agcataaaaa
     3361 gccgccggtg gcggctttag tgcagtgctc tggcgagtag tcctccctca gccgcagtct
     3421 ctgggcctct tcagcttgag cggcggcgag cctgctacac tcgctaagct cctccggcac
     3481 cgcgcacttg ccactgccac tgccgcttcg cgcccgctgc agccgccggc tctgaatcct
     3541 tctggcttcc gcctcagagg agttcttagc ctgtcccgaa ccgtaacccc ggcgagcagt
     3601 ctagaaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     3661 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     3721 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     3781 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     3841 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     3901 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     3961 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     4021 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     4081 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     4141 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     4201 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     4261 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     4321 acaagaagct tagccatggc ttcccgccgg aggtggagga gcaggatgat ggcacgctgc
     4381 ccatgtcttg tgcccaggag agcgggatgg accgtcaccc tgcagcctgt gcttctgcta
     4441 ggatcaatgt gtagaagctt cacatttgta gaggttttac ttgctttaaa aaacctccca
     4501 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     4561 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4621 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaac
     4681 tcgaggcggc cgcgaattcg gtaccggatc caga
//
