LOCUS       sequence-342270-        5164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of Cavin4b-TurboID. Parton lab clone JTA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5164)
  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 5164)
  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-Cavin4b-TurboID
FEATURES             Location/Qualifiers
     source          1..5164
                     /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(1079..1103)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2062..2091
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(2095..2115)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     3'UTR           2140..2265
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(2352..2371)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2353..2371)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2378..2395)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2378..2394)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2387..2409)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2536..2991)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2673..2694)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2885..2904
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3018..3122
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3123..3191,3192..3983)
                     /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(3341..3360)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4154..4742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4643..4662
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4896..4913
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5030..5051
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5066..5071,5072..5089,5090..5096)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5104..5120
                     /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     5109..5131
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5128..5144
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5128..5144
                     /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 accatggctg ataaactggg actggcaggt gtggcggacg agcccagcgc catgaacatc
      181 ttctctctgc tggagagggt gtccgggatc atagacaatg ttcaggcctg ccagcagcgg
      241 atggaggaga ggcagctgga gctggagaac accgtcaaga ccatccaagc agatgtagtc
      301 aagctgacga aggagcacac ggcgaccagt ggcaccgtgg agaaacttct ggagaaaaca
      361 cgcaaggtca gctgccacat caaggacgtc cgcgtgcggg tggagaagca gaacgttcga
      421 gtcaagaagg tggaggaaac ccaagtcgag ctgctgaata agaacaagtt ccgtgtcgtc
      481 atctaccagg gagataatga ggtcccagct gtggctggtc ctaaagcctc tccgaaggga
      541 gcaatgggtg gagacgctgc tgcagaccca gatgcaccag aggctccgct ccctgactcg
      601 gatgaggaat acatggttgt tgaagaagcc gattcttcca ctgctgccaa actaaagaag
      661 accggcctga agcgcatcga aagcctgaag cagaccttct cacgcgagaa catgactaag
      721 accaaagaaa acctgggcac caaggtcaac aagctgggcg agcgcatagt aacagccgaa
      781 cggcgtgaga aaatccgcca gtcaggagaa cgactgaaac agtctggcga gcgcttcaag
      841 gaaaccatca caaaaaccgt accagcaaag ctgaatctga agaaggaaag gacggtggct
      901 gagggtcagg agggggcaga agggaccaca gaaggaacag cacccgtccc accaccgaag
      961 ggtcgtaaac ccagtcctga ggtggcctac acagagccaa cgcagaaaca agaaggtcca
     1021 gatgagggca agggtgagga atcagaagta ccgatgtatg atatgaagca gctttctgac
     1081 ccagctttct tgtacaaagt ggtgaaagac aatacagtcc ccttgaagct gattgccctt
     1141 ctggcaaatg gcgaattcca cagcggggag caactgggcg aaacgttggg catgagtaga
     1201 gcagcgataa acaaacatat acaaacgctg agggattggg gtgtagacgt gttcaccgtt
     1261 ccgggcaagg gctatagttt gcccgaaccg ataccacttc tgaacgccaa acaaatattg
     1321 ggtcaactgg atggaggatc agtagcagtt cttcctgtgg tcgactctac taaccaatat
     1381 ctgctggatc ggatcgggga actgaaatct ggcgatgcat gcatagctga gtaccagcag
     1441 gctggacggg gatctcgagg aaggaaatgg ttttccccgt ttggggccaa cctctatctc
     1501 tctatgttct ggaggttgaa acgtggccct gctgccatcg gcttggggcc tgtcattggt
     1561 attgttatgg cggaggcgtt gcgtaaactt ggagcggata aagtgagggt taaatggcca
     1621 aacgatttgt atcttcaaga ccgcaagttg gctggtatac tggtagaact cgcagggatt
     1681 acaggagatg cggctcagat tgtgatcggt gccggtatta acgtcgcgat gcgtagggta
     1741 gaggagtccg tggtcaatca gggttggatt acgctgcaag aggctggaat caatctcgac
     1801 agaaacacac ttgcagccac gcttattcgg gaattgaggg ctgctcttga actttttgaa
     1861 caggaagggt tggcacctta cctgccgcgc tgggaaaagc tggataattt cataaatcgg
     1921 cccgtcaagc tcataatcgg ggacaaggaa attttcggca tatcacgcgg catcgacaag
     1981 cagggggcac tgctgttgga acaggacgga gtcataaagc cctggatggg tggtgagatt
     2041 agccttcgat ctgcggagaa agagcagaaa ctgataagcg aagaggacct gtaacaactt
     2101 tattatacat agttgctcga gcggatctgg ttaccactaa accagcctca agaacacccg
     2161 aatggagtct ctaagctaca taataccaac ttacacttta caaaatgttg tcccccaaaa
     2221 tgtagccatt cgtatctgct cctaataaaa agaaagtttc ttcacattct aaaaaaaaaa
     2281 aaaaaaaaaa aaaaaaaaan cccccccccc cccctgcagg gccaagtcgg cctctagagg
     2341 tacccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc
     2401 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     2461 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     2521 tgaatggcga atggcaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     2581 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     2641 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2701 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2761 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     2821 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     2881 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     2941 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3001 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3061 tgtatctgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3121 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3181 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3241 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3301 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3361 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3421 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3481 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3541 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3601 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3661 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3721 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     3781 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3841 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3901 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3961 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4021 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4081 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4141 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4201 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4261 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4321 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4381 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4441 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4501 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4561 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4621 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4681 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4741 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4801 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     4861 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     4921 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     4981 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     5041 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     5101 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     5161 tcga
//
