LOCUS       sequence-374528-        5177 bp    DNA     circular SYN 15-APR-2025
DEFINITION  mNeonGreen expression under the control of TRE promotor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5177)
  TITLE     AAV expression plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5177)
  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            "Unpublished"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: pAAV-TRE-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..5177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(82..873,874..942))
                     /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     705..724
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(943..1047)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1115..1133
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1171..1193)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1293..1312
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1329..1784)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1466..1487)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1678..1697
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1859..1999
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1870..1999
                     /label=AAV2 ITR
     polyA_signal    2039..2515
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     2297..2316
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    2547..3135
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2653..2664
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3062..3082
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3160..3870)
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      3606..3615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3867..3876
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3951..3975)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3997..4015
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4034..4052
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4071..4089
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4108..4126
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4145..4163
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4182..4200
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4219..4237
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4256..4274
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     repeat_region   4309..4438
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4500..5088)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4580..4599)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
       61 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
      121 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
      181 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
      241 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
      301 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
      361 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
      421 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
      481 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
      541 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
      601 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
      661 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
      721 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
      781 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
      841 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
      901 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
      961 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     1021 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     1081 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     1141 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     1201 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     1261 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     1321 gcaccataaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     1381 cagctcattt tttaaccaat agaccgaaat cggcaaaatc ccttataaat caaaagaata
     1441 gcccgagata gagttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     1501 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     1561 atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     1621 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     1681 gaagaaagcg aaaggagcgg gcgctaaggc gctggcaagt gtagcggtca cgctgcgcgt
     1741 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg gttgctttga
     1801 cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgcccc
     1861 tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg
     1921 gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac
     1981 tccatcacta ggggttcctg cggccgctcg gtccgcacgt ggttacctac aaaatcagaa
     2041 ggacagggaa gggagcagtg gttcacgcct gtaatcccag caatttggga ggccaaggtg
     2101 ggtagatcac ctgagattag gagttggaga ccagcctggc caatatggtg aaaccccgtc
     2161 tctaccaaaa aaacaaaaat tagctgagcc tggtcatgca tgcctggaat cccaacaact
     2221 cgggaggctg aggcaggaga atcgcttgaa cccaggaggc ggagattgca gtgagccaag
     2281 attgtgccac tgcactccag cttggttccc aatagacccc gcaggcccta caggttgtct
     2341 tcccaacttg ccccttgctc cataccaccc ccctccaccc cataatatta tagaaggaca
     2401 cctagtcaga caaaatgatg caacttaatt ttattaggac aaggctggtg ggcactggag
     2461 tggcaacttc cagggccagg agaggcactg gggaggggtc acagggatgc cacccgtaga
     2521 tctctcgagc agcgctcggt atcgatgcgg ggaggcggcc caaagggaga tccgactcgt
     2581 ctgagggcga aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg
     2641 aaggtccgct ggattgaggg ccgaagggac gtagcagaag gacgtcccgc gcagaatcca
     2701 ggtggcaaca taggcgagca gccaaggaaa ggacgatgat ttccccgaca acaccacgga
     2761 attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag
     2821 ttccgccgtg gcaataggga gggggaaagc gaaagtcccg gaaaggagct gacaggtggt
     2881 ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca gtgcacacca cgccacgttg
     2941 cctgacaacg ggccacaact cctcataaag agacagcaac caggatttat acaaggagga
     3001 gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc
     3061 acatagcgta aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta
     3121 atccagaggt tgattatcga taagcttgat atcgaattct tacttgtaca gctcgtccat
     3181 gcccatcaca tcggtaaagg ccttttgcca ctccttgaag ttgagctcgg tcttggagtg
     3241 cttgagctcc gtcttacgga acacgtacat cggctggttc ttcagatagt tagccgccat
     3301 tggcttggca aaggtgtagg tggtccgcgc agtgctccgg tagcgcttgc catttccagt
     3361 ggtgtaactc cacttaaagg tactgatgat ggttttgtcg ttggggtaag tcttcttcga
     3421 cctgcaccag tccgcagcgg tcagcgagtt ggtcatcaca ggaccgtcag cagggaaacc
     3481 agtccccttc acctgggcct ctcctttgat gtggcttccc tcgtaggtgt agcggtagtt
     3541 aacagtaagg gaggcaccat cttcaaactg cattgtgcga tggacttggt atccggagcc
     3601 atctaccatg gcggcctgga aaggcgacat cccgtcaggg tagggcaggt actgatggaa
     3661 gccatacccg atatgaggga ccagaatcca gggggagaac tggaggtcac ccttggtgga
     3721 cttcaggttt aactcctcat aaccatcatt tggattgccg gtgccctgac ccaccatgtc
     3781 aaagtccaca ccgttgatgg agccaaagat gtgtaactca tgtgtcgctg ggagagaggc
     3841 catgttatcc tcctcgccct tgctcaccat ggtggcggta ccggatccag gctggatcgg
     3901 tcccggtgtc ttctatggag gtcaaaacag cgtggatggc gtctccaggc gatctgacgg
     3961 ttcactaaac gagctctgct tatatagggg gtagcttctc tatcactgat agggagctcg
     4021 agacgtgtcg acttctctat cactgatagg gagacgttct agacgtacct tctctatcac
     4081 tgatagggag ctcgagacgt gtcgacttct ctatcactga tagggagacg ttctagacgt
     4141 accttctcta tcactgatag ggagctcgag acgtgtcgac ttctctatca ctgataggga
     4201 gacgttctag acgtaccttc tctatcactg atagggagct cgagacgtgt cgacttctct
     4261 atcactgata gggagacgtc aggtggcact tgatacgcgt gcggccgcag gaacccctag
     4321 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4381 aggtcgcccg acgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcaggac
     4441 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4501 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4561 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4621 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4681 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4741 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4801 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4861 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4921 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4981 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5041 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5101 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5161 atgagattat caaaaag
//
