LOCUS       sequence-148226-        7715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  acta2:GFP transgenesis construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7715)
  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 7715)
  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: acta2_GFP_pA_pDestTolpA2
FEATURES             Location/Qualifiers
     source          1..7715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(514..525,526..688)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     1061..1077
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1097..1117
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    3582..3606
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(3612..3614,3615..3617,3618..4328)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3612..3614,3615..3617,3618..4328)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3657..3678)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3918..3937)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4265..4286
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(4390..4414)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    4416..4550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4640..4656)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    4672..4806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(5245..5432,5433..5444)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(5478..5494)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5478..5494)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5531..5551)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5531..5549)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5570..5586)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5583..5605)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5594..5610
                     /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)."
     promoter        complement(join(5618..5624,5625..5642,5643..5648))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5663..5684
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5801..5818)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5972..6560)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6052..6071)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6731..7522,7523..7591))
                     /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     7354..7373
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7592..7696)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacgc tggctaagaa
      241 ctcatcagcc tccccggtcc atctacccac gtaccaatgc accaattggc cacaatgacg
      301 gctactacat ggtgccattc cttcctcttt ataggaatgg agactacctc ctgtccaaca
      361 aggctcttgg atacgagtac gcctacctgt tggacccagg tcattgcaca acaccagaaa
      421 tgccctctga tctgcaaaag acgtgaatat ctgttcagac acccatatcc actctgttcc
      481 acacaggtca gaggtttgtc caggagttct tgacagaggt gtaaaaagta ctcaaaaatt
      541 ttactcaagt gaaagtacaa gtacttaggg aaaattttac tcaattaaaa gtaaaagtat
      601 ctggctagaa tcttacttga gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt
      661 attaaggaag taaaagtaaa agcaagaaag aaaactagag attcttgttt aagcttttaa
      721 tctcaaaaaa cattaaatga aatgcataca aggttttatc ctgctttaga actgtttgta
      781 tttaattatc aaactataag acagacaatc taatgccagt acacgctact caaagttgta
      841 aaacctcaga tttaacttca gtagaagctg attctcaaaa ttgttagtgt caagcctagc
      901 tcttttgggg ctgaaaagca atcctgcagt gctgaaaagc ctctcacagg cagccgatgc
      961 gggaagaggt gtattagtct tgatagagag gctgcaaata gcaggaaacg tgagcagaga
     1021 ctccctggtg tctgaaacac aggccagatg ggccctcgag caggaaacag ctatgaccat
     1081 gattacgcca agctatcaac tttgtataga aaagttggcc acctagcttc tctcacctcc
     1141 ttctctttct attttgcaag ctttatctga ttactcttga aactcgctgt gtaaagggtc
     1201 aaagaagtgc tgttgttgtt gttgttgttt tttgcacagg tgtaagctgg ctttaaatat
     1261 tttggctggt tagagtgttt tctgagaaag atttgatcta catttctgta aatgtgattg
     1321 tgaggattga ttttcctaga gacgtgcctt ctgaaagata cattttctac ctatcttcct
     1381 tttctttaaa gaagcactta aagttttaaa gaacaattct gcatggttgt gtctaaagca
     1441 tccatatttt agtatcttta ttgtgatttt atagtaaaat ctctgtgggt gtgagtgttt
     1501 tttaaagaga ggttctgaat ggctgttgaa gcctgtctga tttgttaggt ttttgactaa
     1561 atatggttct tctggaccgc tgaccagtcc cctcttttcc tcacccactc tctctttttt
     1621 cataaactat taggcagaga gagactgaga gagggagaga ccaaagacag acatgcttcc
     1681 ccatcatttc caaattagga agttttccca tggtgctctc agcaagggtc tttaggaaat
     1741 gtgtgtacat atacatgagt gtgtgtgcac atatagaaaa gtacattttt cttcgtaatg
     1801 tattggtatt gttataattg tgttactact aactttttag gatatataac atatatttag
     1861 aaaattacat ttaaaagaca tttaaaatac tttttaaaca ctttaaattt attgatcaat
     1921 gtgtaaaaat taattaacaa attaatccat tatgggtttc ccttattgca ttatgttttg
     1981 ccaattttta gcaataaatg tttttattta tttgttaact ctttaagaac cccaccaaga
     2041 aaaaaaatgt ttggattgca tttcttaact cttaatgtca ctagttacca cactcaatgc
     2101 cttttttaac acatcccata gtttctaaaa tatcagcctc tactaaatgg ttgtaaaatc
     2161 tactaaatct ttattttttg aagcatgaaa taaaatactt cagattctta tttaacatgt
     2221 tttcttgttg ctgttttttt tttttttttt acaataaaac caaaatcgag tgtagtcgta
     2281 caacaggatt atatttgttt gattttcttt gtaaactaac aatgctttgt gtgtgaagca
     2341 ttatttaaag cagtcattat ttaaatgact ttaacattca ttatttaaaa cagttcaaac
     2401 atggtcaacc gtttggtaga ggggcagtca aagggttaat gcttatcaaa ataagcatgt
     2461 tgtttcattt ttacacaaat aatttgtttt actttacaac attgaacata ttttcacccg
     2521 taattgtcat tttcagttat ttgaaacttt tgtacatttt aatttggtgt agatgaacaa
     2581 ctgggactac tcatctttat tattatactt tctcaaaaat gatgtttgtt ttatgtatat
     2641 tatattttat tttacaaaag tttgaattat taaatgtgaa tgatgtcaac gcagagttga
     2701 cgcttgaaat caaaacttct tccttaatgc ttacctggaa aacgaaagat aggcatcaat
     2761 tagtttatga atgaacatag caggaacacc ttgaattcct attattccta tttatagttt
     2821 aatgggtgcg gtccagtctt atcacacaaa actcccatag acataaatgg ttgtttccgg
     2881 tctgatttct gactccgata actgaaaaaa gaaaactttg aaagaacaat tttttcagtt
     2941 tggtcatgct ttactttgat gatccgtttg ttgaatttaa gttcaattgc aactaattct
     3001 cattagatta taagtagact gttaggttgg ggttagggtt agtttaagtt gacatgtact
     3061 tgcaaagttt cttataatca gttaaatgtc tgttgaagta gcagtatcac cagatattaa
     3121 actgacatac agtctactaa tactcaaatg gatcataaaa atcaagggtt accaccagtt
     3181 tatatgaaat gtctccagaa tatgattaag cttaccactg ggagcaaagg tgttctgaaa
     3241 ttaacacaat acattaacaa cacacattgt tgtcatgggc tatgagcgtc tgtgtagcag
     3301 atcaggccca gccctgtgtt tattctcgtg catagtgaag tccccacaat ttctgaggag
     3361 aaaaaaggaa aaggtccatt ggaatgccca gagtaaggag gcaagaggcc gccctggtgc
     3421 tgcccggatt ggagtgtttt ctcccagtca tccctataag gctttagtgg agtgaggcta
     3481 ccgatccgag tgcagcccag gctccttgtt tgggatgtta gagggtgtgg tgagcagaga
     3541 tggaaactac ataagtacac cacaaaccag tgaggtgtac accaagtttg tacaaaaaag
     3601 caggctggac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     3661 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     3721 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     3781 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     3841 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     3901 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     3961 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     4021 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     4081 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     4141 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     4201 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     4261 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     4321 tgtacaaggg cggtggaaga tctgggaatt caaggcctct cgagcctcta gattctgcag
     4381 ccctatagca cccagctttc ttgtacaaag tgggggatcc agacatgata agatacattg
     4441 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     4501 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     4561 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt tccaacttat
     4621 tatacatagt tgataattca ctggccgtcg ttttacggta ccatcgatga tgatccagac
     4681 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     4741 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     4801 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     4861 gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tcctctagat
     4921 cagatctgcg aagatacggc cacgggtgct cttgatcctg tggctgattt tggactgtgc
     4981 tgctcgcagc tgctgatgaa tcacatactt cctccatttt cttccactga ttgactgtta
     5041 taatttccct aatttccagg tcaaggtgct gtgcattgtg gtaatagatg tgacatgacg
     5101 tcacttccaa aggaccaatg aacatgtctg accaatttca tataatgtga aaacgatttt
     5161 cataggcaga ataaataaca tttaaattaa actgggcatc agcgcaattc aattggtttg
     5221 gtaatagcaa gggaaaatag aatgaagtga tctccaaaaa ataagtactt tttgactgta
     5281 aataaaattg taaggagtaa aaagtacttt tttttctaaa aaaatgtaat taagtaaaag
     5341 taaaagtatt gatttttaat tgtactcaag taaagtaaaa atccccaaaa ataatactta
     5401 agtacagtaa tcaagtaaaa ttactcaagt actttacacc tctggttctt gaccccctac
     5461 cttcagcaag cccagcagat ccactagttc tagagcggcc gccaccgcgg tggagctcca
     5521 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     5581 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5641 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5701 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5761 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5821 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5881 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5941 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6001 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6061 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6121 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6181 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6241 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6301 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6361 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     6421 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6481 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6541 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6601 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6661 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6721 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6781 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6841 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6901 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6961 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7021 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7081 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7141 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7201 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7261 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7321 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7381 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7441 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7501 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7561 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7621 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7681 aaataggggt tccgcgcaca tttccccgaa aagtg
//
