LOCUS       sequence-418296-        4449 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Tetraspanin-4a (TSPAN4a) as migrasome marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4449)
  AUTHORS   Jiang D, Jiang Z, Lu D, Wang X, Liang H, Zhang J, Meng Y, Li Y, Wu 
            D, Huang Y, Chen Y, Deng H, Wu Q, Xiong J, Meng A, Yu L
  TITLE     Migrasomes provide regional cues for organ morphogenesis during 
            zebrafish gastrulation.
  JOURNAL   Nat Cell Biol. 2019 Aug;21(8):966-977. doi: 
            10.1038/s41556-019-0358-6. Epub 2019 Aug 1.
  PUBMED    31371827
REFERENCE   2  (bases 1 to 4449)
  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"; doi: 
            "10.1038/s41556-019-0358-6"; journalName: "Nat Cell Biol"; date: 
            "2019-08"; volume: "21"; issue: "8"; pages: "966-977"
COMMENT     Sequence Label: pXTm-zebrafish Tspan4a
FEATURES             Location/Qualifiers
     source          1..4449
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           32..74
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             819..1529
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(964..982)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1232..1251
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1426..1449
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     3'UTR           1563..1688
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(1789..1807)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1790..1807)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1815..1832)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1815..1831)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1824..1846)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2040..2059)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2159..2181
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2219..2237)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2305..2409
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2410..2478,2479..3270)
                     /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(2628..2647)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3441..4029
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3930..3949
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4183..4200
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4317..4338
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4353..4358,4359..4376,4377..4383)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4391..4407
                     /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     4396..4418
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4415..4431
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4415..4431
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 taatacgact cactataggg agacaagctt gcttgttctt tttgcagaag ctcagaataa
       61 acgctcaact ttggcagatc ggtaccgaat tcatggcgga gggctgtctg cgggcgctgc
      121 gatacgggat ggtcttcttc aatctgctct tctggctgtg cggctgtggg attctgggag
      181 ttggagtgtg gctctccatc acgcagggaa acttcgccac actctcgtcc tcgctgccgt
      241 cgctctccgc cgctaacctg ctgatcgccg cgggcaccgt cgtcatggtg atcggctgtc
      301 tgggctgcgt gggtgcggtc aaggagaatc ggccactgct gctcagtttc ttcatcctcc
      361 tgctcctgat cttcctgctg gagatcctct tcatcatcct gttcttctcc tatcaggatc
      421 agattgatct gtatgctcag aatgacctga agaaaggcct gcagctcttc gggactgaag
      481 gaaacatcgg cctgaccaac gcctggagca tcgtgcagac agactttcgc tgctgtggag
      541 tgaccaatca cacagactgg tttcaggtgt ataacacctc tcgtgtgccg gactcctgct
      601 gtctggaata cagcgacaac tgtggcctgg agaaccccgg cacctggtgg acagctccat
      661 gttatgagag agttaaaggt tggctgcagg agaatctggt ggctctgtgg attttcgctc
      721 tgtgtacggc acttacacag atcctgggtc tggtgttctc catgaccatc ttctgtcaca
      781 cagtgaaggc agcagacacc tactatgcgc cagatcttat ggtgagcaag ggcgaggagg
      841 acaacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
      901 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
      961 ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc
     1021 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
     1081 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     1141 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
     1201 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
     1261 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
     1321 agaggctgaa gctgaaggac ggcggccact acgacgccga ggtcaagacc acctacaagg
     1381 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagctg gacatcacct
     1441 cccacaacga ggactacacc atcgtggaac agtacgagcg cgccgagggc cgccactcca
     1501 ccggcggcat ggacgagctg tacaagtaag actagtgact gactaggatc tggttaccac
     1561 taaaccagcc tcaagaacac ccgaatggag tctctaagct acataatacc aacttacact
     1621 ttacaaaatg ttgtccccca aaatgtagcc attcgtatct gctcctaata aaaagaaagt
     1681 ttcttcacat tctaaaaaaa aaaaaaaaaa aaaaaaaaaa aacccccccc ccccccccct
     1741 gcaggtcgac tctagaggat ccccgggcga gctcgaatta attcgtattc tatagtgtca
     1801 cctaaatcca attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     1861 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     1921 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
     1981 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     2041 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     2101 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     2161 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     2221 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     2281 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2341 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2401 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     2461 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     2521 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2581 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     2641 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     2701 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     2761 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     2821 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     2881 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     2941 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3001 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     3061 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     3121 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     3181 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     3241 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     3301 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     3361 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3421 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3481 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3541 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     3601 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3661 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3721 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3781 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3841 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3901 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3961 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4021 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4081 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     4141 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     4201 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     4261 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     4321 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     4381 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     4441 gccaagctc
//
