LOCUS       sequence-342269-        5447 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of CD44b-TurboID. Parton lab clone JTJ.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5447)
  AUTHORS   Xiong Z, Lo HP, McMahon KA, Martel N, Jones A, Hill MM, Parton RG, 
            Hall TE
  TITLE     In vivo proteomic mapping through GFP-directed proximity-dependent 
            biotin labelling in zebrafish.
  JOURNAL   Elife. 2021 Feb 16;10. pii: 64631. doi: 10.7554/eLife.64631.
  PUBMED    33591275
REFERENCE   2  (bases 1 to 5447)
  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: "Elife."; 
            date: "2021-02-16"; volume: "10. pii: 64631. doi"; pages: " 
            10.7554/eLife.64631"
COMMENT     Sequence Label: pT3TS-CD44b-TurboID
FEATURES             Location/Qualifiers
     source          1..5447
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     5'UTR           44..86
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     protein_bind    93..117
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      118..127
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1361..1385)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2344..2373
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(2377..2397)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     3'UTR           2422..2547
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(2635..2654)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2636..2654)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2661..2678)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2661..2677)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2670..2692)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2819..3274)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2956..2977)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3168..3187
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3301..3405
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3406..3474,3475..4266)
                     /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(3624..3643)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4437..5025
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4926..4945
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5179..5196
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5313..5334
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5349..5354,5355..5372,5373..5379)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5387..5403
                     /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     5392..5414
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5411..5427
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5411..5427
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aattaaccct cactaaaggg aacaaaagct ggagctaagc ttgcttgttc tttttgcaga
       61 agctcagaat aaacgctcaa ctttggcaga tctcaagttt gtacaaaaaa gcaggctgcc
      121 accatgggct ggtcatgggt gtgtgttcag acgctccttc agttgacacc cttcctctct
      181 tctctcagtc agtcggactc aatctctcta cactcacaga tgctcttcag gacagagagg
      241 atgatgaaga tgatgatgat gatggcagca cttctgtctg cattgctgtc ctcctccact
      301 gcaaacccaa tcaccggtgt gcgtgagcgg agctgcagct actctggagt gttccatgtg
      361 cagggtaaag agcgctacag cctgacgttt gatgacgcac agagactgtg tgagcgtttg
      421 tccactacat tagccaatct gacacaggtg aagacagcct tcaacagtgg aatggagacc
      481 tgcaggtatg gctggataaa cagctcacag ctggtgattc tccgtaaaac accaaatcta
      541 aactgtgctc gcaaccaaac agacgtcatc ataaagactt cagacgggaa aaaatatgac
      601 gccttatgtt ataatgataa agacaaatca aagaagaact gtacagaggc gattgaccct
      661 gcaaaagtgg acagcagttc atcttcagaa gccgcagttt ctccagagcg catcactcca
      721 cattactcta cagaagcaga ctcctccaca gcagcagttt ctccagagcg catcactcca
      781 cattactcta cagacgcaga ttcctccaca tcagcagttt ctccagcccg cttcactcca
      841 ggcggctcta caggcgcaga ttcctccaca tcagcagaat ctgctctgaa cacgtcagca
      901 gcagcagcag cagcagacgc agagagaaac acaactgagg gaaacacgac acacaccagt
      961 tccccacaca gccacgagga gcccaacggg acacgctcga cccctcagac ctccacagaa
     1021 ctgcagatca gcacgtctgc agcgcatcag gacaccaccg gcccgggccg caggcagaac
     1081 acagtttctc cagagcgcat cactccacac agccctacag aagcagcttc ctccacatca
     1141 gcaggtgctt cttcttcttt aacagactgg ctggtcatat tactgacaat actggcagtt
     1201 cttctcatct cactcatctg ttgtatcgta gtatacagga aaaggctgtt tggacagaag
     1261 cagactctgg tcatcaacag cactgtcgca gcagcagaga acggatcagc ggctacacac
     1321 acacaggaga tggtcacgct gctgaacaca gacacactcc acccagcttt cttgtacaaa
     1381 gtggtgaaag acaatacagt ccccttgaag ctgattgccc ttctggcaaa tggcgaattc
     1441 cacagcgggg agcaactggg cgaaacgttg ggcatgagta gagcagcgat aaacaaacat
     1501 atacaaacgc tgagggattg gggtgtagac gtgttcaccg ttccgggcaa gggctatagt
     1561 ttgcccgaac cgataccact tctgaacgcc aaacaaatat tgggtcaact ggatggagga
     1621 tcagtagcag ttcttcctgt ggtcgactct actaaccaat atctgctgga tcggatcggg
     1681 gaactgaaat ctggcgatgc atgcatagct gagtaccagc aggctggacg gggatctcga
     1741 ggaaggaaat ggttttcccc gtttggggcc aacctctatc tctctatgtt ctggaggttg
     1801 aaacgtggcc ctgctgccat cggcttgggg cctgtcattg gtattgttat ggcggaggcg
     1861 ttgcgtaaac ttggagcgga taaagtgagg gttaaatggc caaacgattt gtatcttcaa
     1921 gaccgcaagt tggctggtat actggtagaa ctcgcaggga ttacaggaga tgcggctcag
     1981 attgtgatcg gtgccggtat taacgtcgcg atgcgtaggg tagaggagtc cgtggtcaat
     2041 cagggttgga ttacgctgca agaggctgga atcaatctcg acagaaacac acttgcagcc
     2101 acgcttattc gggaattgag ggctgctctt gaactttttg aacaggaagg gttggcacct
     2161 tacctgccgc gctgggaaaa gctggataat ttcataaatc ggcccgtcaa gctcataatc
     2221 ggggacaagg aaattttcgg catatcacgc ggcatcgaca agcagggggc actgctgttg
     2281 gaacaggacg gagtcataaa gccctggatg ggtggtgaga ttagccttcg atctgcggag
     2341 aaagagcaga aactgataag cgaagaggac ctgtaacaac tttattatac atagttgctc
     2401 gagcggatct ggttaccact aaaccagcct caagaacacc cgaatggagt ctctaagcta
     2461 cataatacca acttacactt tacaaaatgt tgtcccccaa aatgtagcca ttcgtatctg
     2521 ctcctaataa aaagaaagtt tcttcacatt ctaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2581 annccccccc ccccccctgc agggccaagt cggcctctag aggtacccaa ttcgccctat
     2641 agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     2701 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     2761 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcaa
     2821 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     2881 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     2941 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     3001 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     3061 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3121 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3181 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3241 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     3301 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatct gctcatgaga
     3361 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3421 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3481 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3541 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3601 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     3661 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3721 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3781 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3841 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3901 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     3961 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4021 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4081 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     4141 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     4201 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     4261 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     4321 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4381 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4441 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4501 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4561 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4621 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4681 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4741 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4801 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4861 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4921 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4981 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5041 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5101 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     5161 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     5221 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     5281 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     5341 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     5401 caatttcaca caggaaacag ctatgaccat gattacgcca agctcga
//
