LOCUS       sequence-405350-        6301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cr-dependent (DIO) expression of eGFP-tagged ribosomal subunit that 
            uses the prairie vole RPL10a gene sequence under the human synapsin 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6301)
  AUTHORS   Sadino JM, Bradeen XG, Kelly CJ, Brusman LE, Walker DM, Donaldson ZR
  TITLE     Prolonged partner separation erodes nucleus accumbens 
            transcriptional signatures of pair bonding in male prairie voles.
  JOURNAL   Elife. 2023 Feb 28;12:e80517. doi: 10.7554/eLife.80517.
  PUBMED    36852906
REFERENCE   2  (bases 1 to 6301)
  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: "Elife."; 
            date: "2023-02-28"; volume: "12:e80517. doi"; pages: " 
            10.7554/eLife.80517"
COMMENT     Sequence Label: AAV1-DIO-eGFP-pvRPL10a
FEATURES             Location/Qualifiers
     source          1..6301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(660..693)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(744..777)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(1557..2267,2268..2270,2271..2273))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1557..2267,2268..2270,2271..2273))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1599..1620)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1948..1967
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2207..2228
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2270..2279
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2286..2319
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2370..2403
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2428..3016
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2481..2501)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2899..2910)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3048..3524
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3247..3266)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3564..3704
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3564..3693
                     /label=AAV2 ITR
     rep_origin      3779..4234
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3866..3885)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4076..4097
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4251..4270)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4370..4392
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4430..4448)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4516..4620
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4621..4689,4690..5481)
                     /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(4839..4858)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5652..6240
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6141..6160
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gactccggaa
      661 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
      721 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggt
      781 agagtagcac gaaggtttat tggggcatcc tagtaaagac gctggggctt gcccatggtg
      841 ctcttgatat acaaagcccg gacattttgc cagttcttct tgagcaagga caccaagaaa
      901 ttgacagcca agtggatgtt gtagaccagc tcgtcgtcgg tcatcttcac gtggccgaca
      961 gcgacagcca gacacagcac cttcttcatc tggaacttga tcgtggactt cacctcgtcc
     1021 actttggcca ccatgttttc attgtgtgtc agcagggaag ggaacttgcc agccttgttc
     1081 aggcctgggc ccaggatacg cgggatctgc ttgatcagag actcagaggc cagaaaggcg
     1141 tcatacttct tggccagctt tttgaccaac ttcttgttct tattgagctt cttcagcgcc
     1201 tcgatgtcca tgtgggggat atccacggcc ttggcctcat cgcagtgctg ctggtccccc
     1261 aggacacaca ctgagaactt ggggcgaggg gtggacttga gcctgacggt gcccgagaaa
     1321 cgcttgtcct tctgggggtc gtagttcttc aggctgatct gcagctccac cgtttccaga
     1381 aacttgcggc gcttgcgctg gttcccgtgc aggacctccc gcaccgcctc gtacagggtg
     1441 tcgcgagaga ctttgctgct ggctggagag gctgcgggaa ccggaagaga agacagcgat
     1501 ctgcgcttgc gctcaaagaa ttcgaagctt gagctcgaga tctgagtccg gccggacttg
     1561 tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg
     1621 tgatcgcgct tctcgttggg gtctttgctc agggcggact gggtgctcag gtagtggttg
     1681 tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc
     1741 tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat gccgttcttc
     1801 tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc
     1861 aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg
     1921 tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg
     1981 gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg
     2041 tggtcggggt agcggctgaa gcactgcacg ccgtaggtca gggtggtcac gagggtgggc
     2101 cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgtaggtg
     2161 gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc
     2221 tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac catggtggcg
     2281 ctagcataac ttcgtataaa gtatcctata cgaagttatt tgccttaacc cagaaattat
     2341 cactgttatt ctttagaatg gtgcaaagaa taacttcgta taatgtatgc tatacgaagt
     2401 tatgaattcg atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga
     2461 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     2521 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     2581 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     2641 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     2701 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     2761 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     2821 aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg
     2881 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     2941 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     3001 tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct
     3061 gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc
     3121 ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta
     3181 tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct
     3241 gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct
     3301 ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg
     3361 catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat
     3421 tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat
     3481 tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa
     3541 ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     3601 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     3661 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     3721 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     3781 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3841 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3901 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3961 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     4021 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4081 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     4141 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4201 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     4261 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     4321 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     4381 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     4441 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     4501 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4561 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4621 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4681 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4741 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4801 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4861 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4921 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4981 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5041 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5101 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5161 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5221 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5281 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5341 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5401 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5461 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     5521 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5581 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5641 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5701 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5761 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5821 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5881 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5941 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6001 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6061 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6121 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6181 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6241 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6301 t
//
