LOCUS       sequence-488731-        6334 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of phospho-mutant mouse Ngn2 with HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6334)
  AUTHORS   Lende-Dorn BA, Atkinson JC, Bae Y, Galloway KE
  TITLE     Chemogenetic tuning reveals optimal MAPK signaling for cell-fate 
            programming
  JOURNAL   bioRxiv 2024.12.19.629350
REFERENCE   2  (bases 1 to 6334)
  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: "bioRxiv 
            2024.12.19.629350"
COMMENT     Sequence Label: pMXs-9SA.mNgn2-x3HA
FEATURES             Location/Qualifiers
     source          1..6334
                     /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             1146..1172
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1182..1208
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1218..1244
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(1315..1339)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1356..1944
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1409..1429)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1827..1838)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2124..2593
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2723..2740)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2894..3482)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2974..2993)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3646..4437,4438..4506))
                     /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     4269..4288
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4507..4611)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             4641..5234
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5297..5654
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5713..6129
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6059..6081
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6097..6113
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(6139..6334,1..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 catgggaacc aattcagtcg actggatccg gtactactga attcaagctt
      301 ggatcccaat ggccaccatg ttcgtcaaat ctgagactct ggagttgaag gaggaagagg
      361 aggtactgat gctgctgggc tcggctgccc cggcctcggc gaccctgacc ccgatgtcct
      421 ccagcgcgga cgaggaggag gacgaggagc tgcgccggcc gggctccgcg cgtgggcagc
      481 gtggagcgga agccgggcag ggggtgcagg gcgccccggc gtcgggtgcc gggggttgcc
      541 ggccagggcg gctgctgggc ctgatgcacg agtgcaagcg tcgcccgtcg cgctcacggg
      601 ccgtctcccg aggtgccaag acggcggaga cggtgcagcg catcaagaag acccgcaggc
      661 tcaaggccaa caaccgcgag cgcaaccgca tgcacaacct aaacgccgcg ctggacgcgc
      721 tgcgcgaggt gctgcccacc ttccccgagg atgccaagct cacgaagatc gagacgctgc
      781 gcttcgccca caattacatc tgggcgctca ccgagactct gcgcctggcg gaccactgcg
      841 ccggcgccgg tggcctccag ggggcgctct tcacggaggc ggtgctcctg gccccgggag
      901 ctgcgctcgg cgccagcggg gacgcccctg ccccaccttc ctcctggagc tgcaccaacg
      961 ccccggcgtc atcctccaac tccacggccc catacagctg cactttagcc cccgctgccc
     1021 ccgggtcaga cgtggactac tggcagcccc cacctccgga gaagcatcgt tatgcgcctc
     1081 acctgcccct cgccagggac tgtatcgggt ctggctccgg atcaagtggt ggcagcggat
     1141 caggatatcc atacgatgta ccagactatg cgggatcggg ttacccctac gacgtccccg
     1201 actacgccgg ctccggctac ccctacgacg tccctgatta tgctggttct ggtggtgttg
     1261 aataacaacg ttatcccctg attctgtgga taaccgtatt acgctgatat ctagacccag
     1321 ctttcttgta caaagtggtt cgatagctta tcgataatca acctctggat tacaaaattt
     1381 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     1441 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     1501 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     1561 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     1621 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     1681 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     1741 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     1801 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     1861 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     1921 tctccctttg ggccgcctcc ccgcatcgat accgtcgacg ataaaataaa agattttatt
     1981 tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta
     2041 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga agttcagatc
     2101 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
     2161 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
     2221 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
     2281 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     2341 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     2401 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
     2461 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     2521 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     2581 cgggggtctt tcacatgcag catgtatcaa aattaatttg gttttttttc ttaagtattt
     2641 acattaaatg gccatagttg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2701 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2761 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2821 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2881 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2941 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3001 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3061 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3121 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3181 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3241 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3301 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3361 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3421 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3481 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3541 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttgcggccgg
     3601 ccgcaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3661 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3721 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3781 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3841 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3901 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3961 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4021 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4081 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4141 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4201 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4261 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4321 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4381 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4441 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4501 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4561 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4621 ccgaaaagtg ccacctgcat aatgaaagac cccacctgta ggtttggcaa gctagcttaa
     4681 gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagaaaa gttcagatca
     4741 aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
     4801 tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc aaacaggata
     4861 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg
     4921 tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg
     4981 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
     5041 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct
     5101 ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc
     5161 gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc
     5221 gggggtcttt catttggggg ctcgtccggg atcgggagac ccctgcccag ggaccaccga
     5281 cccaccaccg ggaggtaagc tggccagcaa cttatctgtg tctgtccgat tgtctagtgt
     5341 ctatgactga ttttatgcgc ctgcgtcggt actagttagc taactagctc tgtatctggc
     5401 ggacccgtgg tggaactgac gagttcggaa cacccggccg caaccctggg agacgtccca
     5461 gggacttcgg gggccgtttt tgtggcccga cctgagtcca aaaatcccga tcgttttgga
     5521 ctctttggtg caccccccta ataggaggga tatgtggttc tggtaggaga cgagaaccta
     5581 aaacagttcc cgcctccgtc tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg
     5641 cgtcttgtct gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg
     5701 tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa
     5761 gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc
     5821 ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac
     5881 cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca
     5941 gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc
     6001 aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc gtctctcccc
     6061 cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct cactccttct
     6121 ctaggcgccc ccatatggcc atatgagatc ttatatgggg cacccccgcc ccttgtaaac
     6181 ttccctgacc ctgacatgac aagagttact aacagcccct ctctccaagc tcacttacag
     6241 gctctctact tagtccagca cgaagtctgg agacctctgg cggcagccta ccaagaacaa
     6301 ctggaccgac cggtggtacc tcacccttac cgag
//
