LOCUS       sequence-425821-        4600 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Single stranded AAV eGFP reporter vector with SCP1 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4600)
  TITLE     Cross-species enhancer modeling reveals cell type homologies between
            the mammalian and avian forebrain
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4600)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pSA358_pAAV-SCP1-Intron-eGFP-CS1
FEATURES             Location/Qualifiers
     source          1..4600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   121..250
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          506..735
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     718..737
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      783..792
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(789..791,792..794,795..1508)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(789..791,792..794,795..1505)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(834..855)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1095..1114)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1442..1463
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1581..1601)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   1983..2123
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1983..2112
                     /label=AAV2 ITR
     rep_origin      2198..2653
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2285..2304)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2495..2516
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2670..2689)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2789..2811
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2849..2867)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2935..3039
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3040..3108,3109..3900)
                     /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(3258..3277)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4071..4600,1..59)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4560..4579
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
       61 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
      121 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
      181 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      241 aggggttcct gcggccgcag atgctgcaca cttgtggact aagtttgttc gcatcccctt
      301 ctccaacccc ctcagtacat caccctgggg gaacagggtc cacttgctcc tgggccctgc
      361 gtatgagtct gagctcccta gggtacttat ataagggggt gggggcgcgt tcgtcctcag
      421 tcgcgatcga acactcgagc cgagcagacg tgcctacgga ccggggcccg aattaattcg
      481 ctgtctgcga gggccagctg ttggggtgag tactccctct caaaagcggg catgacttct
      541 gcgctaagat tgtcagtttc caaaaacgag gaggatttga tattcacctg gcccgcggtg
      601 atgcctttga gggtggccgc gtccatctgg tcagaaaaga caatcttttt gttgtcaagc
      661 ttgaggtgtg gcaggcttga gatctggcca tacacttgag tgacaatgac atccactttg
      721 cctttctctc cacaggtgtc cactcccagg tgtcgaattc aagctgctag ctagtaccgg
      781 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      841 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      901 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      961 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     1021 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1081 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1141 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1201 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     1261 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     1321 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1381 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     1441 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1501 acaagtaaat cgaattccgc tcgagataat caacctctgg attacaaaat ttgtgaaaga
     1561 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     1621 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     1681 tggttagttc ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     1741 gctcggctgt tgggcactga caattccgtg gtgctttaag gccggtccta gcaagtttat
     1801 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatc tagctttatt tgtgaaattt
     1861 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1921 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcggcc
     1981 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     2041 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     2101 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     2161 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     2221 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     2281 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2341 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2401 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2461 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2521 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     2581 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2641 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     2701 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     2761 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     2821 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     2881 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     2941 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3001 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3061 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3121 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3181 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3241 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     3301 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3361 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3421 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3481 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3541 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     3601 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3661 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3721 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3781 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3841 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3901 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3961 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4021 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4081 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4141 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4201 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4261 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4321 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4381 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4441 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4501 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4561 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
//
