LOCUS       sequence-308775-        6249 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Independent expression of mOrange and human Kvbeta 1 under human 
            synapsin 1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6249)
  AUTHORS   Cho IH, Panzera LC, Chin M, Alpizar SA, Olveda GE, Hill RA, Hoppa MB
  TITLE     The potassium channel subunit K(v)beta1 serves as a major control 
            point for synaptic facilitation.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Nov 24;117(47):29937-29947. doi: 
            10.1073/pnas.2000790117. Epub 2020 Nov 9.
  PUBMED    33168717
REFERENCE   2  (bases 1 to 6249)
  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"; doi: 
            "10.1073/pnas.2000790117"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2020-11-24- 24"; volume: "117"; issue: "47"; pages: 
            "29937-29947"
COMMENT     Sequence Label: mOrange-P2A-hKvb1
FEATURES             Location/Qualifiers
     source          1..6249
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   55..184
                     /label=AAV2 ITR
     rep_origin      complement(421..1009)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(501..520)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1180..1971,1972..2040))
                     /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     1803..1822
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2041..2145)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2171..2626)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2308..2329)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2520..2539
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2698..2827
                     /label=AAV2 ITR
     promoter        2909..3356
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      3385..3394
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3391..4098
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEYNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             4108..4164
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     misc_feature    5384..5972
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5437..5457)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5855..5866)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5975..5991)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5976..5992)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    6021..6142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6032..6051)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6086..6105
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 tcctagagca tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac
       61 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
      121 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
      181 gcagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
      241 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
      301 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
      361 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
      421 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
      481 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      541 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      601 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      661 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      721 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      781 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      841 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      901 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      961 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     1021 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     1081 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     1141 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     1201 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     1261 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     1321 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     1381 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1441 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     1501 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1561 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1621 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1681 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1741 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1801 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1861 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1921 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1981 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     2041 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     2101 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     2161 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     2221 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     2281 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     2341 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     2401 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     2461 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     2521 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     2581 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
     2641 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     2701 cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt
     2761 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
     2821 ggttccttgt agttaatgat taacccgcca tgctacttat ctacgtagcc atgctctagg
     2881 aagatctctg cagagggccc tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc
     2941 ggggtggggg tgcctacctg acgaccgacc ccgacccact ggacaagcac ccaaccccca
     3001 ttccccaaat tgcgcatccc ctatcagaga gggggagggg aaacaggatg cggcgaggcg
     3061 cgtgcgcact gccagcttca gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc
     3121 gccaccgccg cctcagcact gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc
     3181 cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg
     3241 cgaggcgcga gatagggggg cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag
     3301 cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcga
     3361 attcaagctg ctagcaagga tcccgccacc atggtgagca agggcgagga gtataacatg
     3421 gccatcatca aggagttcat gcgcttcaag gtgcgcatgg agggctccgt gaacggccac
     3481 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggctttca gaccgctaag
     3541 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcatttc
     3601 acctacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttcaagctg
     3661 tccttccccg agggcttcaa gtgggagcgc gtgatgaact acgaggacgg cggcgtggtg
     3721 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
     3781 ggcaccaact tcccctccga cggccccgtg atgcagaaga agaccatggg ctgggaggcc
     3841 tcctccgagc ggatgtaccc cgaggacggt gccctgaagg gcaagatcaa gatgaggctg
     3901 aagctgaagg acggcggcca ctacacctcc gaggtcaaga ccacctacaa ggccaagaag
     3961 cccgtgcagc tgcccggcgc ctacatcgtc gacatcaagt tggacatcac ctcccacaac
     4021 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     4081 atggacgagc tgtacaaggg aagcggagct actaacttca gcctgctgaa gcaggctgga
     4141 gacgtggagg agaaccctgg acctatgcaa gtctccatag cctgcacaga gcacaatttg
     4201 aagagtcgga atggtgagga ccgacttctg agcaagcaga gctccaccgc ccccaatgtg
     4261 gtgaacgcag cccgggccaa attccgcacg gtcgctatca tcgcgcgcag cctggggacg
     4321 ttcacgcctc agcatcacat ttctctcaaa gagtccaccg caaagcagac tggcatgaaa
     4381 tataggaatc ttggaaaatc aggactcaga gtttcttgct tgggtcttgg aacatgggtg
     4441 acatttggag gtcaaatttc agatgaggtt gctgaacggc tgatgaccat cgcctatgaa
     4501 agtggtgtta acctctttga tactgccgaa gtctatgctg ctggaaaggc tgaagtgatt
     4561 ctggggagca tcatcaagaa gaaaggctgg aggaggtcca gtctggtcat aacaaccaaa
     4621 ctctactggg gtggaaaagc tgaaacagaa agagggctgt caagaaagca tattattgaa
     4681 ggattgaagg gctccctcca gaggctgcag ctcgagtatg tggatgtggt ctttgcaaat
     4741 cgaccggaca gtaacactcc catggaagaa attgtccgag ccatgacaca tgtgataaac
     4801 caaggcatgg cgatgtactg gggcacctcg agatggagtg ctatggagat catggaagcc
     4861 tattctgtag caagacagtt caatatgatc ccaccggtct gtgaacaagc tgagtaccat
     4921 cttttccaga gagagaaagt ggaggtccag ctgccagagc tctaccacaa aataggtgtt
     4981 ggcgcaatga catggtctcc acttgcctgt ggaatcatct caggaaaata cggaaacggg
     5041 gtgcctgaaa gttccagggc ttcactgaag tgctaccagt ggttgaaaga aagaattgta
     5101 agtgaagaag ggagaaaaca gcaaaacaag ctaaaagacc tttccccaat tgcggagcgt
     5161 ctgggatgca cactacctca gctagctgtt gcgtggtgcc tgagaaatga aggtgtgagt
     5221 tctgtgctcc tgggatcatc cactcctgaa caactcattg aaaaccttgg tgccattcag
     5281 gttctcccaa agatgacatc acatgtggta aatgagattg ataacatact gcgcaacaag
     5341 ccctacagca agaaggacta tagatcataa gaagcttatc gataatcaac ctctggatta
     5401 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     5461 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     5521 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     5581 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     5641 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     5701 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     5761 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     5821 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     5881 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     5941 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gacccgggcg
     6001 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     6061 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     6121 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     6181 gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
     6241 taaggatct
//
