LOCUS       sequence-255888-        6448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV vector that expresses rat NK1R-IRES-EGFP under the CMV-IE 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6448)
  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 6448)
  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: pOTTC946 - pAAV CMV-IE NK1R-IRES-EGFP
FEATURES             Location/Qualifiers
     source          1..6448
                     /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"
     regulatory      739..748
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(1997..2571,2572..2574,2575..2583)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2164..2181)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2391..2410
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2584..2586,2587..2589,2590..3303)
                     /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(2584..2586,2587..2589,2590..3300)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2629..2650)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2890..2909)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3237..3258
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3429..3550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3466..3485)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3520..3539
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3711..3851
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3711..3840
                     /label=AAV2 ITR
     rep_origin      3926..4381
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4013..4032)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4223..4244
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4398..4417)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4517..4539
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4577..4595)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4663..4767
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4768..4836,4837..5628)
                     /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(4986..5005)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5799..6387
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6288..6307
                     /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 caccatggat aacgtccttc ctatggactc tgatctcttc
      781 cccaacatct ccaccaacac ctctgagtct aaccagttcg tgcagcctac ctggcaaatc
      841 gttctttggg cggctgccta cacggtcatc gtggtgactt ccgtggtggg caacgtagtg
      901 gtgatatgga tcattttggc ccacaagaga atgaggacag tgaccaatta tttcctggtg
      961 aacctggcct tcgctgaggc ctgcatggct gcattcaata cggtggtgaa cttcacctac
     1021 gcagtccaca atgtgtggta ctacggcctc ttctattgca agtttcacaa cttcttcccc
     1081 atcgctgctc tcttcgccag tatctactcc atgacagccg tggccttcga cagatacatg
     1141 gccatcatcc accctctcca gccccggctc tcagccacgg ctaccaaagt ggtcatcttt
     1201 gtcatctggg tcctggctct cctgctggcc tttccacaag gctactactc caccacagag
     1261 accatgccca gcagagtcgt gtgcatgatc gagtggccgg agcatcccaa caggacttat
     1321 gagaaagcgt accacatctg cgtgactgta ctgatctact tcctgcctct actggtgatc
     1381 ggctacgcgt acactgtggt ggggattaca ctgtgggcca gtgagatccc cggcgactcc
     1441 tctgaccgct accatgagca agtctctgcc aaacgcaagg tggtcaaaat gatgatcgtg
     1501 gttgtgtgta ccttcgccat ctgctggctg cccttccacg tcttcttcct cctgccctac
     1561 atcaacccag atctctacct gaagaagttc atccagcagg tctacctggc cagcatgtgg
     1621 ctggccatga gttctaccat gtacaacccc atcatctact gttgcctcaa cgacaggttc
     1681 cgtctgggct tcaagcacgc cttccgctgc tgccccttca tcagtgcagg tgattatgag
     1741 gggctggaaa tgaaatccac ccgatacctc cagacacaga gcagcgtgta caaggtcagc
     1801 cgcctggaga ccaccatctc cactgtggtg ggagcccatg aagaggagcc cgaggaaggc
     1861 cccaaggcca caccctcatc cttggatctc acctccaacg gctcctctcg cagcaactcc
     1921 aagaccatga cagaaagctc cagcttctac tctaacatgc tggcctaggt cgacggtacc
     1981 gcgggcccgg gatccgcccc tctccctccc ccccccctaa cgttactggc cgaagccgct
     2041 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     2101 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     2161 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     2221 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     2281 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     2341 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     2401 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     2461 atctggggcc tcggtacaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     2521 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca
     2581 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     2641 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     2701 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     2761 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     2821 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2881 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2941 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     3001 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     3061 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     3121 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     3181 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     3241 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     3301 taaagcggcc gcgactctag aggatcataa tcagccatac cacatttgta gaggttttac
     3361 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3421 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3481 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3541 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3601 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3661 aaagaataga ccgagatagg gttgagtgca cgtgcggacc gagcggccgc aggaacccct
     3721 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     3781 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     3841 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3901 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3961 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     4021 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4081 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4141 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4201 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     4261 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     4321 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     4381 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     4441 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4501 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4561 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4621 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4681 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4741 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4801 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4861 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4921 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4981 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5041 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5101 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5161 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5221 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5281 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5341 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5401 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5461 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5521 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5581 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5641 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5701 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5761 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5821 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5881 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5941 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6001 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6061 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6121 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6181 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6241 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6301 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6361 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6421 gccttttgct ggccttttgc tcacatgt
//
