LOCUS       sequence-255887-        5239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV vector that expresses IRES EGFP under the CMV-IE promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5239)
  AUTHORS   Nelson BS, Fulenwider HD, Nennig SE, Smith BM, Sequeira MK, 
            Chimberoff SH, Richie CT, Cheng K, Rice KC, Harvey BK, Heilig M, 
            Schank JR
  TITLE     Escalated Alcohol Self-Administration and Sensitivity to 
            Yohimbine-Induced Reinstatement in Alcohol Preferring Rats: 
            Potential Role of Neurokinin-1 Receptors in the Amygdala.
  JOURNAL   Neuroscience. 2019 Jun 23;413:77-85. doi: 
            10.1016/j.neuroscience.2019.06.023.
  PUBMED    31242442
REFERENCE   2  (bases 1 to 5239)
  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: 
            "Neuroscience."; date: "2019-06-23"; volume: "413:77-85. doi"; 
            pages: " 10.1016/j.neuroscience.2019.06.023"
COMMENT     Sequence Label: pOTTC385 - pAAV CMV-IE IRES EGFP
FEATURES             Location/Qualifiers
     source          1..5239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        205..508
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        509..712
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     663..683
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    join(788..1362,1363..1365,1366..1374)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(955..972)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1182..1201
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1375..1377,1378..1380,1381..2094)
                     /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(1375..1377,1378..1380,1381..2091)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1420..1441)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1681..1700)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2028..2049
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2220..2341
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2257..2276)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2311..2330
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   2502..2642
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2502..2631
                     /label=AAV2 ITR
     rep_origin      2717..3172
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2804..2823)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3014..3035
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3189..3208)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3308..3330
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3368..3386)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3454..3558
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3559..3627,3628..4419)
                     /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(3777..3796)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4590..5178
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5079..5098
                     /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 gcgttagtta ttaatagtaa tcaattacgg ggtcattagt
      181 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      241 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      301 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      361 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      421 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      481 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      541 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      601 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      661 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
      721 gaaccgtcag atccgctagc gctaccggtg aattctgcag tcgacggtac cgcgggcccg
      781 ggatccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag
      841 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
      901 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
      961 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     1021 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     1081 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     1141 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     1201 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     1261 ctcggtacac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     1321 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtg
     1381 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1441 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     1501 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1561 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     1621 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1681 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1741 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1801 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     1861 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     1921 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1981 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     2041 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc
     2101 cgcgactcta gaggatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     2161 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     2221 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     2281 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     2341 aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     2401 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     2461 accgagatag ggttgagtgc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     2521 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     2581 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     2641 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     2701 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2761 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     2821 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2881 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     2941 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3001 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     3061 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     3121 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     3181 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     3241 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     3301 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     3361 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     3421 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3481 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3541 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3601 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3661 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3721 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3781 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     3841 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3901 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3961 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4021 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4081 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     4141 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4201 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4261 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4321 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4381 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4441 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4501 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4561 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4621 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4681 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4741 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4801 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4861 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4921 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4981 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5041 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5101 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5161 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5221 tggccttttg ctcacatgt
//
