LOCUS       sequence-148229-        5107 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  acta2 promoter for Gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5107)
  AUTHORS   Whitesell TR, Kennedy RM, Carter AD, Rollins EL, Georgijevic S, 
            Santoro MM, Childs SJ
  TITLE     An alpha-smooth muscle actin (acta2/alphasma) zebrafish transgenic 
            line marking vascular mural cells and visceral smooth muscle cells.
  JOURNAL   PLoS One. 2014 Mar 3;9(3):e90590. doi: 10.1371/journal.pone.0090590.
            eCollection 2014.
  PUBMED    24594685
REFERENCE   2  (bases 1 to 5107)
  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: "PLoS One.";
            date: "2014-03-3"; volume: "9(3):e90590. doi"; pages: " 
            10.1371/journal.pone.0090590. eCollection 2014"
COMMENT     Sequence Label: p5E-acta2
FEATURES             Location/Qualifiers
     source          1..5107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    592..687
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    3153..3276
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3348..3367)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3349..3367)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3372..3388)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3372..3388)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3490..3509)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3501..4310
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3568..3587)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4457..5045
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4946..4965
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccgc gttaacgcta ccatggagct ccaataatga
      601 ttttattttg actgatagtg acctgttcgt tgcaacaaat tgataagcaa tgctttttta
      661 taatgccaac tttgtataga aaagttggcc acctagcttc tctcacctcc ttctctttct
      721 attttgcaag ctttatctga ttactcttga aactcgctgt gtaaagggtc aaagaagtgc
      781 tgttgttgtt gttgttgttt tttgcacagg tgtaagctgg ctttaaatat tttggctggt
      841 tagagtgttt tctgagaaag atttgatcta catttctgta aatgtgattg tgaggattga
      901 ttttcctaga gacgtgcctt ctgaaagata cattttctac ctatcttcct tttctttaaa
      961 gaagcactta aagttttaaa gaacaattct gcatggttgt gtctaaagca tccatatttt
     1021 agtatcttta ttgtgatttt atagtaaaat ctctgtgggt gtgagtgttt tttaaagaga
     1081 ggttctgaat ggctgttgaa gcctgtctga tttgttaggt ttttgactaa atatggttct
     1141 tctggaccgc tgaccagtcc cctcttttcc tcacccactc tctctttttt cataaactat
     1201 taggcagaga gagactgaga gagggagaga ccaaagacag acatgcttcc ccatcatttc
     1261 caaattagga agttttccca tggtgctctc agcaagggtc tttaggaaat gtgtgtacat
     1321 atacatgagt gtgtgtgcac atatagaaaa gtacattttt cttcgtaatg tattggtatt
     1381 gttataattg tgttactact aactttttag gatatataac atatatttag aaaattacat
     1441 ttaaaagaca tttaaaatac tttttaaaca ctttaaattt attgatcaat gtgtaaaaat
     1501 taattaacaa attaatccat tatgggtttc ccttattgca ttatgttttg ccaattttta
     1561 gcaataaatg tttttattta tttgttaact ctttaagaac cccaccaaga aaaaaaatgt
     1621 ttggattgca tttcttaact cttaatgtca ctagttacca cactcaatgc cttttttaac
     1681 acatcccata gtttctaaaa tatcagcctc tactaaatgg ttgtaaaatc tactaaatct
     1741 ttattttttg aagcatgaaa taaaatactt cagattctta tttaacatgt tttcttgttg
     1801 ctgttttttt tttttttttt acaataaaac caaaatcgag tgtagtcgta caacaggatt
     1861 atatttgttt gattttcttt gtaaactaac aatgctttgt gtgtgaagca ttatttaaag
     1921 cagtcattat ttaaatgact ttaacattca ttatttaaaa cagttcaaac atggtcaacc
     1981 gtttggtaga ggggcagtca aagggttaat gcttatcaaa ataagcatgt tgtttcattt
     2041 ttacacaaat aatttgtttt actttacaac attgaacata ttttcacccg taattgtcat
     2101 tttcagttat ttgaaacttt tgtacatttt aatttggtgt agatgaacaa ctgggactac
     2161 tcatctttat tattatactt tctcaaaaat gatgtttgtt ttatgtatat tatattttat
     2221 tttacaaaag tttgaattat taaatgtgaa tgatgtcaac gcagagttga cgcttgaaat
     2281 caaaacttct tccttaatgc ttacctggaa aacgaaagat aggcatcaat tagtttatga
     2341 atgaacatag caggaacacc ttgaattcct attattccta tttatagttt aatgggtgcg
     2401 gtccagtctt atcacacaaa actcccatag acataaatgg ttgtttccgg tctgatttct
     2461 gactccgata actgaaaaaa gaaaactttg aaagaacaat tttttcagtt tggtcatgct
     2521 ttactttgat gatccgtttg ttgaatttaa gttcaattgc aactaattct cattagatta
     2581 taagtagact gttaggttgg ggttagggtt agtttaagtt gacatgtact tgcaaagttt
     2641 cttataatca gttaaatgtc tgttgaagta gcagtatcac cagatattaa actgacatac
     2701 agtctactaa tactcaaatg gatcataaaa atcaagggtt accaccagtt tatatgaaat
     2761 gtctccagaa tatgattaag cttaccactg ggagcaaagg tgttctgaaa ttaacacaat
     2821 acattaacaa cacacattgt tgtcatgggc tatgagcgtc tgtgtagcag atcaggccca
     2881 gccctgtgtt tattctcgtg catagtgaag tccccacaat ttctgaggag aaaaaaggaa
     2941 aaggtccatt ggaatgccca gagtaaggag gcaagaggcc gccctggtgc tgcccggatt
     3001 ggagtgtttt ctcccagtca tccctataag gctttagtgg agtgaggcta ccgatccgag
     3061 tgcagcccag gctccttgtt tgggatgtta gagggtgtgg tgagcagaga tggaaactac
     3121 ataagtacac cacaaaccag tgaggtgtac accaagtttg tacaaaaaag ttgaacgaga
     3181 aacgtaaaat gatataaata tcaatatatt aaattagatt ttgcataaaa aacagactac
     3241 ataatactgt aaaacacaac atatgcagtc actatgaacc aactacttag atggtattag
     3301 tgacctgtag aattcgagct ctagagctgc agggcggccg cgatatcccc tatagtgagt
     3361 cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat
     3421 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
     3481 acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa
     3541 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
     3601 caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac
     3661 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga
     3721 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt
     3781 tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt
     3841 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg
     3901 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa
     3961 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
     4021 aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc
     4081 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
     4141 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
     4201 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     4261 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg
     4321 ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat
     4381 gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa
     4441 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4501 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4561 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     4621 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4681 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4741 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4801 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg
     4861 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4921 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4981 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5041 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5101 acatgtt
//
