LOCUS       sequence-146928-        5104 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 5104)
  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 5104)
  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..5104
                     /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    593..688
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3345..3364)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3346..3364)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3369..3385)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3369..3385)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3487..3506)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3498..4307
                     /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(3565..3584)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4454..5042
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4943..4962
                     /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 ccaaataatg
      601 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca atgctttttt
      661 ataatgccaa ctttgtatag aaaagttgaa tatagaaaag ttggccacct agcttctctc
      721 acctccttct ctttctattt tgcaagcttt atctgattac tcttgaaact cgctgtgtaa
      781 agggtcaaag aagtgctgtt gttgttgttg ttgttttttg cacaggtgta agctggcttt
      841 aaatattttg gctggttaga gtgttttctg agaaagattt gatctacatt tctgtaaatg
      901 tgattgtgag gattgatttt cctagagacg tgccttctga aagatacatt ttctacctat
      961 cttccttttc tttaaagaag cacttaaagt tttaaagaac aattctgcat ggttgtgtct
     1021 aaagcatcca tattttagta tctttattgt gattttatag taaaatctct gtgggtgtga
     1081 gtgtttttta aagagaggtt ctgaatggct gttgaagcct gtctgatttg ttaggttttt
     1141 gactaaatat ggttcttctg gaccgctgac cagtcccctc ttttcctcac ccactctctc
     1201 ttttttcata aactattagg cagagagaga ctgagagagg gagagaccaa agacagacat
     1261 gcttccccat catttccaaa ttaggaagtt ttcccatggt gctctcagca agggtcttta
     1321 ggaaatgtgt gtacatatac atgagtgtgt gtgcacatat agaaaagtac atttttcttc
     1381 gtaatgtatt ggtattgtta taattgtgtt actactaact ttttaggata tataacatat
     1441 atttagaaaa ttacatttaa aagacattta aaatactttt taaacacttt aaatttattg
     1501 atcaatgtgt aaaaattaat taacaaatta atccattatg ggtttccctt attgcattat
     1561 gttttgccaa tttttagcaa taaatgtttt tatttatttg ttaactcttt aagaacccca
     1621 ccaagaaaaa aaatgtttgg attgcatttc ttaactctta atgtcactag ttaccacact
     1681 caatgccttt tttaacacat cccatagttt ctaaaatatc agcctctact aaatggttgt
     1741 aaaatctact aaatctttat tttttgaagc atgaaataaa atacttcaga ttcttattta
     1801 acatgttttc ttgttgctgt tttttttttt ttttttacaa taaaaccaaa atcgagtgta
     1861 gtcgtacaac aggattatat ttgtttgatt ttctttgtaa actaacaatg ctttgtgtgt
     1921 gaagcattat ttaaagcagt cattatttaa atgactttaa cattcattat ttaaaacagt
     1981 tcaaacatgg tcaaccgttt ggtagagggg cagtcaaagg gttaatgctt atcaaaataa
     2041 gcatgttgtt tcatttttac acaaataatt tgttttactt tacaacattg aacatatttt
     2101 cacccgtaat tgtcattttc agttatttga aacttttgta cattttaatt tggtgtagat
     2161 gaacaactgg gactactcat ctttattatt atactttctc aaaaatgatg tttgttttat
     2221 gtatattata ttttatttta caaaagtttg aattattaaa tgtgaatgat gtcaacgcag
     2281 agttgacgct tgaaatcaaa acttcttcct taatgcttac ctggaaaacg aaagataggc
     2341 atcaattagt ttatgaatga acatagcagg aacaccttga attcctatta ttcctattta
     2401 tagtttaatg ggtgcggtcc agtcttatca cacaaaactc ccatagacat aaatggttgt
     2461 ttccggtctg atttctgact ccgataactg aaaaaagaaa actttgaaag aacaattttt
     2521 tcagtttggt catgctttac tttgatgatc cgtttgttga atttaagttc aattgcaact
     2581 aattctcatt agattataag tagactgtta ggttggggtt agggttagtt taagttgaca
     2641 tgtacttgca aagtttctta taatcagtta aatgtctgtt gaagtagcag tatcaccaga
     2701 tattaaactg acatacagtc tactaatact caaatggatc ataaaaatca agggttacca
     2761 ccagtttata tgaaatgtct ccagaatatg attaagctta ccactgggag caaaggtgtt
     2821 ctgaaattaa cacaatacat taacaacaca cattgttgtc atgggctatg agcgtctgtg
     2881 tagcagatca ggcccagccc tgtgtttatt ctcgtgcata gtgaagtccc cacaatttct
     2941 gaggagaaaa aaggaaaagg tccattggaa tgcccagagt aaggaggcaa gaggccgccc
     3001 tggtgctgcc cggattggag tgttttctcc cagtcatccc tataaggctt tagtggagtg
     3061 aggctaccga tccgagtgca gcccaggctc cttgtttggg atgttagagg gtgtggtgag
     3121 cagagatgga aactacataa gtacaccaca aaccagtgag gtgtacactg aacgagaaac
     3181 gtaaaatgat ataaatatca atatattaaa ttagattttg cataaaaaac agactacata
     3241 atactgtaaa acacaacata tgcagtcact atgaatcaac tacttagatg gtattagtga
     3301 cctgtagaat tcgagctcta gagctgcagg gcggccgcga tatcccctat agtgagtcgt
     3361 attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt
     3421 acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca
     3481 gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt
     3541 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     3601 gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     3661 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     3721 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     3781 tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag
     3841 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     3901 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga
     3961 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     4021 aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg
     4081 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     4141 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     4201 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     4261 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta
     4321 attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac
     4381 caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga
     4441 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4501 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4561 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     4621 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4681 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4741 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4801 tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga gaaagcgcca
     4861 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4921 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4981 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5041 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5101 tgtt
//
