LOCUS       sequence-334625-        4640 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encoding partial sense strand of the mouse Tbxas1 gene for in situ 
            hybridization..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4640)
  AUTHORS   Konishi Y, Ichise H, Watabe T, Oki C, Tsukiji S, Hamazaki Y, 
            Murakawa Y, Takaori-Kondo A, Terai K, Matsuda M
  TITLE     Intravital imaging identifies the VEGF-TXA2 axis as a critical 
            promoter of PGE2 secretion from tumor cells and immune evasion.
  JOURNAL   Cancer Res. 2021 May 25. pii: 0008-5472.CAN-20-4245. doi: 
            10.1158/0008-5472.CAN-20-4245.
  PUBMED    34035084
REFERENCE   2  (bases 1 to 4640)
  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: "Cancer Res.
            2021 May 25. pii: 0008-5472.CAN-20-4245. doi: 
            10.1158/0008-5472.CAN-20-4245."
COMMENT     Sequence Label: Topo-Tbxas1_ISH_S
FEATURES             Location/Qualifiers
     source          1..4640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1014..1033)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1015..1033)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1040..1057)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1040..1056)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1049..1071)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1194..1496
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1375..1394
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1845..2639
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1845..2639
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1899..1918)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2509..2528
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      2839..2848
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3185..3563)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     3510..3529
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3871..4459
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4360..4379
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4613..4630
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga cyatgrkyrm ryyrrgcymt
      241 ytagatgcat gctcgagcgg ccgccagtgt gatggatatc tgcagaattc gccctttgct
      301 tccccatgaa gaggtccacc tctttcagaa gcctctcctg gcagtcaggg tgggtggcca
      361 gcaggtatgt gatgaaggac agcgtgtttg tgatgacctc atggcccgca atgaggaaga
      421 ggaaggcctg gcccacaatt tcatccacag tgaaaggctt agatgtggag gtaggatggc
      481 acctttgggg tggttccttt gtgcactcag aagaggacag ggattctggg accatgtcaa
      541 agccttccac gcccacagag ttcatggagt gctgggcatc cagcaccatc tgcaggaagt
      601 ctctccgcct ctcttctgct gcttgctggt cccgtaaggc aatcacattc ctaatgagtg
      661 tgttaaaaaa gccattcagt tcatctcggt tcttattggg cagaatccgg gccaatggga
      721 ccattatgga tggaaatgat aagattaata ccaggagagg cctggggata cagaaggtgg
      781 aagcacgccg gcagtgttgc acaaagggat cttctggaga attctgggag tccacctggg
      841 tgccaaaggc cacactggcc accacatcta tggtgtaaca gcagtaacac ctctggatgt
      901 tgaatgcgtc cctggatgct gcatagcgtt ttaagtgagc cacgagaagt tcacaggctt
      961 ggctgatgag asrykksmks mmgskcgary wyncatctag agggcccaat tcgccctata
     1021 gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     1081 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     1141 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctatacgta cggcagttta
     1201 aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata
     1261 ttattgacac gccggggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag
     1321 ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga
     1381 tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg gctgatctca
     1441 gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatgt
     1501 caggcatgag attatcaaaa aggatcttca cctagatcct tttcacgtag aaagccagtc
     1561 cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg acaagggaaa
     1621 acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga tagctagact
     1681 gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc tctggtaagg
     1741 ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca
     1801 ggggatcaag ctctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     1861 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     1921 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     1981 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     2041 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     2101 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     2161 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     2221 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     2281 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     2341 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     2401 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     2461 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     2521 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     2581 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgaa
     2641 ttattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     2701 cacaccgcat caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2761 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2821 taatagcacg tgaggagggc caccatggcc aagttgacca gtgccgttcc ggtgctcacc
     2881 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
     2941 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
     3001 gtccaggact gstgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3061 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3121 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3181 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3241 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3301 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3361 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3421 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3481 gagttgctct tgcccggcgt caayacggga taataccgcg ccacatagca gaactttaaa
     3541 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3601 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcmk scgagcagcc
     3661 gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga
     3721 ggagcaggac tgacacgtgc taaaacttca tttttaattt aaaaggatct aggtgaagat
     3781 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3841 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3901 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3961 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4021 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4081 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4141 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4201 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4261 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4321 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4381 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4441 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4501 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4561 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     4621 agtgagcgag gaagcggaag
//
