LOCUS       sequence-410040-        7633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Halo-tagged mouse SOX17FNV in mammalian cells, for 
            single-molecule tracking..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7633)
  AUTHORS   Weng M, Hu H, Graus MS, Tan DS, Gao Y, Ren S, Ho DHH, Langer J, 
            Holzner M, Huang Y, Ling GS, Lai CSW, Francois M, Jauch R
  TITLE     An engineered Sox17 induces somatic to neural stem cell fate 
            transitions independently from pluripotency reprogramming.
  JOURNAL   Sci Adv. 2023 Aug 25;9(34):eadh2501. doi: 10.1126/sciadv.adh2501. 
            Epub 2023 Aug 23.
  PUBMED    37611093
REFERENCE   2  (bases 1 to 7633)
  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: "Sci Adv."; 
            date: "2023-08-25"; volume: "9(34):eadh2501. doi"; pages: " 
            10.1126/sciadv.adh2501. Epub 2023 Aug 23"
COMMENT     Sequence Label: Halo-tagged Sox17FNV
FEATURES             Location/Qualifiers
     source          1..7633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(2..26)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    64..287
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      333..761
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(420..439)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     630..651
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(770..790)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        775..1104
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      955..1090
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1017..1036
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1171..1965
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1225..1244)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1835..1854
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2141..2262
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2178..2197)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2232..2251
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2311..2327)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2311..2327)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2324..2346)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2335..2351
                     /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)."
     promoter        complement(join(2359..2365,2366..2383,2384..2389))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2404..2425
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2542..2559)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2713..3301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2793..2812)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3472..4263,4264..4332))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEDRFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4095..4114
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4333..4437)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4512..4531)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4703..5082
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5083..5286
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5237..5257
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5331..5350
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5331..5349
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5379..6269
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             6282..6302
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     protein_bind    6324..6348
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 aacccagctt tcttgtacaa agtggttcga tctagaatgg ctagcctcga gtttaaacct
       61 cgactgtgct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
      121 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
      181 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
      241 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
      301 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
      361 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
      421 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
      481 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
      541 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
      601 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
      661 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
      721 gttaaaaaat gagctgattt aacaaaaaat taacgcgaat taatgctgtg gaatgtgtgt
      781 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
      841 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
      901 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
      961 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     1021 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     1081 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     1141 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     1201 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     1261 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     1321 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     1381 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     1441 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     1501 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     1561 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     1621 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     1681 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     1741 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     1801 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     1861 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     1921 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     1981 tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     2041 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     2101 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     2161 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2221 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     2281 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     2341 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     2401 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2461 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2521 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2581 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2641 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2701 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2761 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2821 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2881 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2941 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3001 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3061 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3121 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3181 gaagccagtt accttcggaa aaggagttgg tagctcttga tccggcaaac aaaccaccgc
     3241 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3301 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3361 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3421 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3481 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3541 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3601 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3661 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3721 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3781 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3841 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3901 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3961 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4021 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4081 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4141 aaaacggtct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4201 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4261 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4321 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtcc
     4381 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4441 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     4501 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     4561 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     4621 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
     4681 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
     4741 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     4801 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     4861 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     4921 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     4981 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     5041 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     5101 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     5161 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     5221 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     5281 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
     5341 cactataggg agacccaagc tggctagtta agcttaccat ggcagaaatc ggtactggct
     5401 ttccattcga cccccattat gtggaagtcc tgggcgagcg catgcactac gtcgatgttg
     5461 gtccgcgcga tggcacccct gtgctgttcc tgcacggtaa cccgacctcc tcctacgtgt
     5521 ggcgcaacat catcccgcat gttgcaccga cccatcgctg cattgctcca gacctgatcg
     5581 gtatgggcaa atccgacaaa ccagacctgg gttatttctt cgacgaccac gtccgcttca
     5641 tggatgcctt catcgaagcc ctgggtctgg aagaggtcgt cctggtcatt cacgactggg
     5701 gctccgctct gggtttccac tgggccaagc gcaatccaga gcgcgtcaaa ggtattgcat
     5761 ttatggagtt catccgccct atcccgacct gggacgaatg gccagaattt gcccgcgaga
     5821 ccttccaggc cttccgcacc accgacgtcg gccgcaagct gatcatcgat cagaacgttt
     5881 ttatcgaggg tacgctgccg atgggtgtcg tccgcccgct gactgaagtc gagatggacc
     5941 attaccgcga gccgttcctg aatcctgttg accgcgagcc actgtggcgc ttcccaaacg
     6001 agctgccaat cgccggtgag ccagcgaaca tcgtcgcgct ggtcgaagaa tacatggact
     6061 ggctgcacca gtcccctgtc ccgaagctgc tgttctgggg caccccaggc gttctgatcc
     6121 caccggccga agccgctcgc ctggccaaaa gcctgcctaa ctgcaaggct gtggacatcg
     6181 gcccgggtct gaatctgctg caagaagaca acccggacct gatcggcagc gagatcgcgc
     6241 gctggctgtc gacgctcgag atttccggcg agccaaccac tgaggatctg tactttcaga
     6301 gcgataacgc gatcgccgaa ggaacaagtt tgtacaaaaa agcaggcttg aaggaattcg
     6361 gtaccatgag cagcccggat gcgggatacg ccagtgacga ccagagccag ccccggagcg
     6421 cgcagcccgc ggtgatggca gggttgggcc cctgtccctg ggccgagtcc ctgagccccc
     6481 tcggggatgt aaaggtgaaa ggcgaggtgg tggcgagtag cggggcgcca gccgggacgt
     6541 cgggccgagc caaagcggag tctcgcatcc ggcggccgat gaacgccttt atggtgtggg
     6601 ccaaagacga acgcaagcgg ttggcacagc agaacccaga tctgcacaac gcagagctaa
     6661 gcaagatgct aggcaagtct tggaaggcgt tgacctttgc agagaagcgg cccttcaatg
     6721 aagaggccgt gcggctgcgc gtgcagcata tgcaggacca ccccaactac aagtaccggc
     6781 cgcggcggcg caagcaggtg aagcgcatga agcgggtgga gggaggcttc ctgcacgctc
     6841 tcgtcgagcc ccaggccggc gcgcttggtc ccgagggcgg ccgcgtggcc atggatggcc
     6901 tgggtctgcc tttcccggag ccgggctatc cggccggtcc tccgctgatg tctccgcaca
     6961 tgggccccca ctatcgggac tgccagggac tgggcgctcc cgcgctcgac ggctaccctc
     7021 tgcccactcc ggacacatcc ccgctggatg gcgtggagca ggacccggct ttctttgcag
     7081 ccccgctgcc aggggactgc ccggcggccg gcacctacac ttacgctcca gtctcggact
     7141 atgcagtgtc cgtagagccg cccgctggcc ccatgcgagt ggggccggac ccctcgggcc
     7201 ctgcgatgcc ggggatcctg gcgcccccca gcgctctgca cctgtactac ggcgcgatgg
     7261 gctcgcccgc cgcaagtgcg gggcgcggtt tccacgcgca accccagcag ccgctgcaac
     7321 cgcaggcacc gcccccgcca ccgcagcagc agcacccagc gcacggcccc gggcaacctt
     7381 cgccccctcc cgaggctctg ccctgccggg atggcacgga atccaaccag cccactgagc
     7441 tcctagggga ggtggaccgc acggaattcg aacagtatct gccctttgtg tataagcccg
     7501 agatgggtct tccctaccag ggacacgact gcggagtgaa cctctcagac agccacggag
     7561 ccatttcctc cgtggtgtcc gacgctagct cagcggtcta ctattgcaac taccccgaca
     7621 tttaagcggc cgc
//
