LOCUS       sequence-462991-        6703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of jRCaMP1b calcium reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6703)
  AUTHORS   Wang NB, Adewumi HO, Lende-Dorn BA, Beitz AM, O'Shea TM, Galloway KE
  TITLE     Compact transcription factor cassettes generate functional, 
            engraftable motor neurons by direct conversion.
  JOURNAL   Cell Syst. 2025 Mar 10:101206. doi: 10.1016/j.cels.2025.101206.
  PUBMED    40086435
REFERENCE   2  (bases 1 to 6703)
  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: "Cell Syst. 
            2025 Mar 10:101206. doi: 10.1016/j.cels.2025.101206."
COMMENT     Sequence Label: pMXs-jRCaMP1b
FEATURES             Location/Qualifiers
     source          1..6703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     117..133
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     protein_bind    222..246
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             355..372
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             376..408
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(412..420,421..435)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     protein_bind    complement(1684..1708)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1725..2313
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1778..1798)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2196..2207)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2493..2962
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3092..3109)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3263..3851)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3343..3362)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4015..4806,4807..4875))
                     /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     4638..4657
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4876..4980)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5010..5603
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5666..6023
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6082..6498
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6428..6450
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6466..6482
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(6508..6703,1..179)
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
ORIGIN
        1 tcggcgacac agtgtgggtc cgccgacacc agactaagaa cctagaacct cgctggaaag
       61 gaccttacac agtcctgctg accaccccca ccgccctcaa agtagacggc atcgcagctt
      121 ggatacacgc cgcccacgtg aaggctgccg accccggggg tggaccatcc tctagactgc
      181 cggatctagc tagttaatcg acggtatcga taagctatca aacaagtttg tacaaaaaag
      241 caggctccac caggaaccaa ttcagtcgac tggatccggt acgccaccat gctgcagaac
      301 gagcttgctc ttaagttggc tggacttgat attaacaaga ctggaggagg ttctcatcat
      361 catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg ggatctgtac
      421 gacgatgacg ataaggatct cgcaacaatg gtcgactcat cgcgacgtaa gtggaataag
      481 tggggtcacg cagtcagagc tataggtcgg ctgagctcag cgaacaacac cgaaatgatg
      541 tacccagcgg atggtggtct gcgtggttac actcacatgg cgctgaaagt tgatggcggc
      601 ggtcacctgt cctgttcttt cgtgaccacc taccgctcca aaaagactgt cggcaacatt
      661 aagatgcctg ccattcatta cgtcagccac cgtctggagc gcctggagga gagcgataac
      721 gaaatgtttg tcgtacagcg tgaacacgca gttgccaagt ttgtgggcct gggtggtggc
      781 ggcggtaccg gagggagcat gaactccctg atcaaggaga acatgcgtat gaaagtggtt
      841 ctggaaggct ccgtaaacgg ccaccagttc aaatgcactg gtgaaggcga aggcaacccg
      901 tatatgggca cccagactat gcgtatcaaa gtgatcgagg gtggtccgct gccgtttgcg
      961 ttcgacatcc tggcgacgtc ctttatgtat ggctcccgta ccttcatcaa atatccgaaa
     1021 ggcatcccgg atttctttaa gcagtccttc ccggaaggtt ttacctggga acgtgtgacc
     1081 cgttacgaag acggcggcgt aattaccgtt atgcaagaca cgtctctgga ggatggctgc
     1141 ctggtgtatc acgtgcaggt tcgcggtgtg aacttcccga gcaatggtgc tgtaatgcaa
     1201 aagaaaacca aaggttggga gcctacggac tcccaactga ctgaagagca gatcgcagaa
     1261 tttaaagagg ctttctccct atttgacaag gacggggatg ggacaataac aaccaaggag
     1321 atggggacgg tgatgcggtc tctggggcag aaccccacag aagcagagct gcaggacatg
     1381 atcaatgaag tagatgccga cggtgacggc acaatcgact tccctgagtt cctgattatg
     1441 atggcaggca aaatgaaata cacagacagt gaagaagaaa ttagagaagc gttcggcgtg
     1501 tttgataagg atggcaatgg ctacatcagt gcagcagagc ttcgccacgt gatgacaaac
     1561 cttggagaga agttaacaga tgaagaggtt gatgaaatga tcagggaagc agacagcgat
     1621 ggggatggtc aggtaaacta cgaagagttt gtacaaatga tgacagcgaa gtgagatatc
     1681 tagacccagc tttcttgtac aaagtggttc gatagcttat cgataatcaa cctctggatt
     1741 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     1801 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     1861 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     1921 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     1981 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     2041 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     2101 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     2161 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     2221 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     2281 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacga taaaataaaa
     2341 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     2401 gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa
     2461 gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt
     2521 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc
     2581 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     2641 ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     2701 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     2761 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg
     2821 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
     2881 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     2941 acccgtcagc gggggtcttt cacatgcagc atgtatcaaa attaatttgg ttttttttct
     3001 taagtattta cattaaatgg ccatagttgc attaatgaat cggccaacgc gcggggagag
     3061 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3121 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3181 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3241 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3301 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3361 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3421 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3481 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3541 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3601 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3661 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3721 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3781 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3841 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3901 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3961 tgcggccggc cgcaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4021 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4081 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4141 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4201 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4261 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4321 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4381 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4441 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4501 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4561 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4621 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4681 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4741 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4801 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4861 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4921 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4981 cacatttccc cgaaaagtgc cacctgcata atgaaagacc ccacctgtag gtttggcaag
     5041 ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag aatagaaaag
     5101 ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
     5161 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca
     5221 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     5281 cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     5341 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     5401 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc
     5461 gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca
     5521 gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta
     5581 cccgtcagcg ggggtctttc atttgggggc tcgtccggga tcgggagacc cctgcccagg
     5641 gaccaccgac ccaccaccgg gaggtaagct ggccagcaac ttatctgtgt ctgtccgatt
     5701 gtctagtgtc tatgactgat tttatgcgcc tgcgtcggta ctagttagct aactagctct
     5761 gtatctggcg gacccgtggt ggaactgacg agttcggaac acccggccgc aaccctggga
     5821 gacgtcccag ggacttcggg ggccgttttt gtggcccgac ctgagtccaa aaatcccgat
     5881 cgttttggac tctttggtgc acccccctaa taggagggat atgtggttct ggtaggagac
     5941 gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg ggaccgaagc
     6001 cgcgccgcgc gtcttgtctg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt
     6061 ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt
     6121 cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt
     6181 tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc
     6241 acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat
     6301 ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct
     6361 ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg
     6421 tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc
     6481 actccttctc taggcgcccc catatggcca tatgagatct tatatggggc acccccgccc
     6541 cttgtaaact tccctgaccc tgacatgaca agagttacta acagcccctc tctccaagct
     6601 cacttacagg ctctctactt agtccagcac gaagtctgga gacctctggc ggcagcctac
     6661 caagaacaac tggaccgacc ggtggtacct cacccttacc gag
//
