LOCUS       sequence-298436-        6559 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 6559)
  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 6559)
  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..6559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   259..388
                     /label=AAV2 ITR
     promoter        470..917
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      948..957
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             954..1661
                     /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             1671..1727
                     /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    2947..3535
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3000..3020)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3418..3429)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3538..3554)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3539..3555)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3584..3705
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3595..3614)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3649..3668
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3867..3996
                     /label=AAV2 ITR
     primer_bind     complement(4015..4032)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4015..4031)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4024..4046)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4173..4628
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4260..4279)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4470..4491
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4654..4758
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4759..4827,4828..5619)
                     /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(4977..4996)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5790..6378
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6279..6298
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccagattta
      241 attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      301 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      361 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      421 tctacgtagc catgctctag gaagatctct gcagagggcc ctgcgtatga gtgcaagtgg
      481 gttttaggac caggatgagg cggggtgggg gtgcctacct gacgaccgac cccgacccac
      541 tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag agggggaggg
      601 gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt
      661 cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac
      721 tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg
      781 gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc gaccatctgc
      841 gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt
      901 cgtgcctgag agcgcagtcg agaattcaag ctgctagcaa ggatcccgcc accatggtga
      961 gcaagggcga ggagtataac atggccatca tcaaggagtt catgcgcttc aaggtgcgca
     1021 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     1081 acgagggctt tcagaccgct aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     1141 gggacatcct gtcccctcat ttcacctacg gctccaaggc ctacgtgaag caccccgccg
     1201 acatccccga ctacttcaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     1261 actacgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     1321 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtgatgcaga
     1381 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggtgccctga
     1441 agggcaagat caagatgagg ctgaagctga aggacggcgg ccactacacc tccgaggtca
     1501 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacatc gtcgacatca
     1561 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     1621 agggccgcca ctccaccggc ggcatggacg agctgtacaa gggaagcgga gctactaact
     1681 tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctatg caagtctcca
     1741 tagcctgcac agagcacaat ttgaagagtc ggaatggtga ggaccgactt ctgagcaagc
     1801 agagctccac cgcccccaat gtggtgaacg cagcccgggc caaattccgc acggtcgcta
     1861 tcatcgcgcg cagcctgggg acgttcacgc ctcagcatca catttctctc aaagagtcca
     1921 ccgcaaagca gactggcatg aaatatagga atcttggaaa atcaggactc agagtttctt
     1981 gcttgggtct tggaacatgg gtgacatttg gaggtcaaat ttcagatgag gttgctgaac
     2041 ggctgatgac catcgcctat gaaagtggtg ttaacctctt tgatactgcc gaagtctatg
     2101 ctgctggaaa ggctgaagtg attctgggga gcatcatcaa gaagaaaggc tggaggaggt
     2161 ccagtctggt cataacaacc aaactctact ggggtggaaa agctgaaaca gaaagagggc
     2221 tgtcaagaaa gcatattatt gaaggattga agggctccct ccagaggctg cagctcgagt
     2281 atgtggatgt ggtctttgca aatcgaccgg acagtaacac tcccatggaa gaaattgtcc
     2341 gagccatgac acatgtgata aaccaaggca tggcgatgta ctggggcacc tcgagatgga
     2401 gtgctatgga gatcatggaa gcctattctg tagcaagaca gttcaatatg atcccaccgg
     2461 tctgtgaaca agctgagtac catcttttcc agagagagaa agtggaggtc cagctgccag
     2521 agctctacca caaaataggt gttggcgcaa tgacatggtc tccacttgcc tgtggaatca
     2581 tctcaggaaa atacggaaac ggggtgcctg aaagttccag ggcttcactg aagtgctacc
     2641 agtggttgaa agaaagaatt gtaagtgaag aagggagaaa acagcaaaac aagctaaaag
     2701 acctttcccc aattgcggag cgtctgggat gcacactacc tcagctagct gttgcgtggt
     2761 gcctgagaaa tgaaggtgtg agttctgtgc tcctgggatc atccactcct gaacaactca
     2821 ttgaaaacct tggtgccatt caggttctcc caaagatgac atcacatgtg gtaaatgaga
     2881 ttgataacat actgcgcaac aagccctaca gcaagaagga ctatagatca taagaagctt
     2941 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     3001 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     3061 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     3121 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     3181 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     3241 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     3301 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     3361 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     3421 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     3481 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga
     3541 taccgtcgac ctcgacccgg gcggccgctt cgagcagaca tgataagata cattgatgag
     3601 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     3661 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     3721 attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac
     3781 ctctacaaat gtggtaaaat cgataaggat cttcctagag catggctacg tagataagta
     3841 gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc
     3901 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     3961 tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt aattaaccta attcactggc
     4021 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     4081 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     4141 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     4201 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4261 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4321 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4381 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4441 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4501 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4561 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     4621 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4681 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4741 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4801 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4861 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4921 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4981 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5041 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5101 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5161 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5221 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5281 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5341 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5401 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5461 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5521 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5581 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5641 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5701 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5761 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5821 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5881 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5941 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6001 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6061 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6121 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6181 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6241 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6301 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6361 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6421 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6481 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     6541 gtgagcgagg aagcggaag
//
