LOCUS       sequence-355960-        7451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses iCre-venus. The promoter is a fragment of the hdc gene 
            promoter that gives pan neuronal expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7451)
  AUTHORS   Miracca G, Anuncibay-Soto B, Tossell K, Yustos R, Vyssotski AL, 
            Franks NP, Wisden W
  TITLE     NMDA Receptors in the Lateral Preoptic Hypothalamus Are Essential 
            for Sustaining NREM and REM Sleep.
  JOURNAL   J Neurosci. 2022 May 31. pii: JNEUROSCI.0350-21.2022. doi: 
            10.1523/JNEUROSCI.0350-21.2022.
  PUBMED    35649726
REFERENCE   2  (bases 1 to 7451)
  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: "J Neurosci.
            2022 May 31. pii: JNEUROSCI.0350-21.2022. doi: 
            10.1523/JNEUROSCI.0350-21.2022."
COMMENT     Sequence Label: pAAV-iCre-2A-venus
FEATURES             Location/Qualifiers
     source          1..7451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        88..106
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     88..105
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     repeat_region   127..256
                     /label=AAV2 ITR
     CDS             1543..1563
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2578..2631
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(2647..2649,2650..2652,2653..3366)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2692..2713)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2953..2972)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3300..3321
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3397..3985
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3450..3470)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3868..3879)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4012..4029)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4018..4242
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(4452..4781)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4466..4601
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4766..4786
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(5027..5615)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5107..5126)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5786..6577,6578..6646))
                     /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     6409..6428
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6647..6751)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6819..6837
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6875..6897)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6997..7016
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7230..7249
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gccagatgct acacaattag gcttgtacat attgtcgtta gaacgcggct acaattaata
       61 cataacctta tgtatcatac acatacgatt taggtgacac tatagaatac acggaattaa
      121 ttctagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
      181 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
      241 atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca
      301 tgctctaggt accgaccacc ccccatgaag tctgttgtgt aagtacaggt aatgctttag
      361 acactgtttt tctcagattg tgttcaagct cttgggtgca ggcatccgca gtccagagcc
      421 tctgaggaac aaagggacct cgatataaag aacaaaaagc tctcggtatt cgggaaaact
      481 ccgaaggatt tttgttgttt tgtggcagtg ttggggttca ggacaaacac accaccccag
      541 aatatgattt caggagaaac tgccatcccc gcccccacta gtacgcttcc tttgtgttat
      601 ctcgagaagc cctggacgta ggaggagttc tcaaaagctg ttctttcgta agagatggtc
      661 aggaagaaac gttgattagg aaaagtactc gactcaggaa gaggaggggt acagataact
      721 gctgtcacca gaaagatttc catctgacaa agtggcaatt cttccccctt acgaatgtgt
      781 gtcccgcact ctgctgcacc caggctccgc tctgttctcc agctctgctt gctgtagaag
      841 cgtccatcca ctaaagggac ttccagattt attggcactg aacaggtttt tcctccgtta
      901 gcttaaacag tagtcgaggc aaagaaccca gaacaagtgc tatgtagtaa tggataaaaa
      961 gaatccttca tatcctgttg ttgatctgtc ctgagctcag cctggctagc tatgggcttc
     1021 tctgagccgg ctctgccctc acctcataga gaagccaggg caggagcaac tccaccctct
     1081 gtgctactta ttgtgaatgc ccttggccta tttctggcct ctgagctagg agctttattc
     1141 aaggctgggg ctctatggtc tgaatgcagg ggcagggatg atagagggct cttttacctt
     1201 gtctatccag agctgcttgt aggcttggga ggctaggtgc cccaactggt ctaaaaacac
     1261 catgttctct gattctgttg atcaaagttc aaatcagaaa ggacggggac aagagaggct
     1321 gaaattaagc ctgaaggaag ggactttatg ggggcggggc taaaggaggg gaatagggaa
     1381 ctccttaaat aagagatgca ctggctgcca gagagtgcac agcacagaca aaggcgacca
     1441 agaagccctg ttgacagctg ccttccagcc tcctctgtct gtctgccagg aggagcaatc
     1501 caaggaattc agagctcacg tcgaagcttg tccaccatgg tgcccaagaa gaagaggaaa
     1561 gtctccaacc tgctgactgt gcaccaaaac ctgcctgccc tccctgtgga tgccacctct
     1621 gatgaagtca ggaagaacct gatggacatg ttcagggaca ggcaggcctt ctctgaacac
     1681 acctggaaga tgctcctgtc tgtgtgcaga tcctgggctg cctggtgcaa gctgaacaac
     1741 aggaaatggt tccctgctga acctgaggat gtgagggact acctcctgta cctgcaagcc
     1801 agaggcctgg ctgtgaagac catccaacag cacctgggcc agctcaacat gctgcacagg
     1861 agatctggcc tgcctcgccc ttctgactcc aatgctgtgt ccctggtgat gaggagaatc
     1921 agaaaggaga atgtggatgc tggggagaga gccaagcagg ccctggcctt tgaacgcact
     1981 gactttgacc aagtcagatc cctgatggag aactctgaca gatgccagga catcaggaac
     2041 ctggccttcc tgggcattgc ctacaacacc ctgctgcgca ttgccgaaat tgccagaatc
     2101 agagtgaagg acatctcccg caccgatggt gggagaatgc tgatccacat tggcaggacc
     2161 aagaccctgg tgtccacagc tggtgtggag aaggccctgt ccctgggggt taccaagctg
     2221 gtggagagat ggatctctgt gtctggtgtg gctgatgacc ccaacaacta cctgttctgc
     2281 cgggtcagaa agaatggtgt ggctgcccct tctgccacct cccaactgtc cacccgggcc
     2341 ctggaaggga tctttgaggc cacccaccgc ctgatctatg gtgccaagga tgactctggg
     2401 cagagatacc tggcctggtc tggccactct gccagagtgg gtgctgccag ggacatggcc
     2461 agggctggtg tgtccatccc tgaaatcatg caggctggtg gctggaccaa tgtgaacatt
     2521 gtgatgaact acatcagaaa cctggactct gagactgggg ccatggtgag gctgctcgag
     2581 ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tgcaccggga
     2641 tccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2701 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     2761 acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg
     2821 cccaccctcg tgaccaccct gggctacggc ctgcagtgct tcgcccgcta ccccgaccac
     2881 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     2941 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     3001 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     3061 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag
     3121 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     3181 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3241 aaccactacc tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     3301 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     3361 aagtaaagcg gccgcaagct ttaacagctt atcgataatc aacctctgga ttacaaaatt
     3421 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3481 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3541 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     3601 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     3661 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     3721 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     3781 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     3841 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     3901 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     3961 atctcccttt gggccgcctc cccgcatcga taccgtcgac tcgctgatca gcctcgactg
     4021 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     4081 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     4141 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     4201 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     4261 ccagctgggg ctcgactaga gcatggctac gtagataagt agcatggcgg gttaatcatt
     4321 aactacaagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     4381 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc
     4441 gcgcagagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat
     4501 agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg
     4561 ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg
     4621 actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     4681 ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
     4741 gctggggagc ctggggactt tccacaccct aactgacaca cattccacag ctgcattaat
     4801 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4861 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4921 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4981 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5041 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5101 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5161 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5221 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5281 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5341 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5401 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5461 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5521 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5581 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5641 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5701 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5761 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5821 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5881 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5941 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6001 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6061 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6121 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6181 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6241 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6301 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6361 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6421 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6481 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6541 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6601 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6661 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6721 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6781 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     6841 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     6901 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     6961 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     7021 gagtgcacca ttcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
     7081 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
     7141 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     7201 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     7261 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat ctggctagcg
     7321 atgaccctgc tgattggttc gctgaccatt tccgggtgcg ggacggcgtt accagaaact
     7381 cagaaggttc gtccaaccaa accgactctg acggcagttt acgagagaga tgatagggtc
     7441 tgcttcagta a
//
