LOCUS       Exported                7665 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  acta2:mCherry transgenesis construct.
ACCESSION   .
VERSION     .
KEYWORDS    acta2_mCherry_pA_pDestTolpA2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7665)
  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 7665)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7665
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(160..181)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1077..1093
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1113..1133
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    3598..3622
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3628..4338
                     /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(3773..3791)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4041..4060
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4235..4258
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(4340..4364)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    4366..4500
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4590..4606)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    4622..4756
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5428..5444)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5428..5444)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5481..5501)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5481..5499)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5520..5536)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5533..5555)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5544..5560
                     /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(5568..5598)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5613..5634
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5751..5768)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5922..6510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6002..6021)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6681..7541)
                     /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     7304..7323
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7542..7646)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 actamchrry adsttaccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
       61 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
      121 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
      181 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
      241 ccacgctggc taagaactca tcagcctccc cggtccatct acccacgtac caatgcacca
      301 attggccaca atgacggcta ctacatggtg ccattccttc ctctttatag gaatggagac
      361 tacctcctgt ccaacaaggc tcttggatac gagtacgcct acctgttgga cccaggtcat
      421 tgcacaacac cagaaatgcc ctctgatctg caaaagacgt gaatatctgt tcagacaccc
      481 atatccactc tgttccacac aggtcagagg tttgtccagg agttcttgac agaggtgtaa
      541 aaagtactca aaaattttac tcaagtgaaa gtacaagtac ttagggaaaa ttttactcaa
      601 ttaaaagtaa aagtatctgg ctagaatctt acttgagtaa aagtaaaaaa gtactccatt
      661 aaaattgtac ttgagtatta aggaagtaaa agtaaaagca agaaagaaaa ctagagattc
      721 ttgtttaagc ttttaatctc aaaaaacatt aaatgaaatg catacaaggt tttatcctgc
      781 tttagaactg tttgtattta attatcaaac tataagacag acaatctaat gccagtacac
      841 gctactcaaa gttgtaaaac ctcagattta acttcagtag aagctgattc tcaaaattgt
      901 tagtgtcaag cctagctctt ttggggctga aaagcaatcc tgcagtgctg aaaagcctct
      961 cacaggcagc cgatgcggga agaggtgtat tagtcttgat agagaggctg caaatagcag
     1021 gaaacgtgag cagagactcc ctggtgtctg aaacacaggc cagatgggcc ctcgagcagg
     1081 aaacagctat gaccatgatt acgccaagct atcaactttg tatagaaaag ttggccacct
     1141 agcttctctc acctccttct ctttctattt tgcaagcttt atctgattac tcttgaaact
     1201 cgctgtgtaa agggtcaaag aagtgctgtt gttgttgttg ttgttttttg cacaggtgta
     1261 agctggcttt aaatattttg gctggttaga gtgttttctg agaaagattt gatctacatt
     1321 tctgtaaatg tgattgtgag gattgatttt cctagagacg tgccttctga aagatacatt
     1381 ttctacctat cttccttttc tttaaagaag cacttaaagt tttaaagaac aattctgcat
     1441 ggttgtgtct aaagcatcca tattttagta tctttattgt gattttatag taaaatctct
     1501 gtgggtgtga gtgtttttta aagagaggtt ctgaatggct gttgaagcct gtctgatttg
     1561 ttaggttttt gactaaatat ggttcttctg gaccgctgac cagtcccctc ttttcctcac
     1621 ccactctctc ttttttcata aactattagg cagagagaga ctgagagagg gagagaccaa
     1681 agacagacat gcttccccat catttccaaa ttaggaagtt ttcccatggt gctctcagca
     1741 agggtcttta ggaaatgtgt gtacatatac atgagtgtgt gtgcacatat agaaaagtac
     1801 atttttcttc gtaatgtatt ggtattgtta taattgtgtt actactaact ttttaggata
     1861 tataacatat atttagaaaa ttacatttaa aagacattta aaatactttt taaacacttt
     1921 aaatttattg atcaatgtgt aaaaattaat taacaaatta atccattatg ggtttccctt
     1981 attgcattat gttttgccaa tttttagcaa taaatgtttt tatttatttg ttaactcttt
     2041 aagaacccca ccaagaaaaa aaatgtttgg attgcatttc ttaactctta atgtcactag
     2101 ttaccacact caatgccttt tttaacacat cccatagttt ctaaaatatc agcctctact
     2161 aaatggttgt aaaatctact aaatctttat tttttgaagc atgaaataaa atacttcaga
     2221 ttcttattta acatgttttc ttgttgctgt tttttttttt ttttttacaa taaaaccaaa
     2281 atcgagtgta gtcgtacaac aggattatat ttgtttgatt ttctttgtaa actaacaatg
     2341 ctttgtgtgt gaagcattat ttaaagcagt cattatttaa atgactttaa cattcattat
     2401 ttaaaacagt tcaaacatgg tcaaccgttt ggtagagggg cagtcaaagg gttaatgctt
     2461 atcaaaataa gcatgttgtt tcatttttac acaaataatt tgttttactt tacaacattg
     2521 aacatatttt cacccgtaat tgtcattttc agttatttga aacttttgta cattttaatt
     2581 tggtgtagat gaacaactgg gactactcat ctttattatt atactttctc aaaaatgatg
     2641 tttgttttat gtatattata ttttatttta caaaagtttg aattattaaa tgtgaatgat
     2701 gtcaacgcag agttgacgct tgaaatcaaa acttcttcct taatgcttac ctggaaaacg
     2761 aaagataggc atcaattagt ttatgaatga acatagcagg aacaccttga attcctatta
     2821 ttcctattta tagtttaatg ggtgcggtcc agtcttatca cacaaaactc ccatagacat
     2881 aaatggttgt ttccggtctg atttctgact ccgataactg aaaaaagaaa actttgaaag
     2941 aacaattttt tcagtttggt catgctttac tttgatgatc cgtttgttga atttaagttc
     3001 aattgcaact aattctcatt agattataag tagactgtta ggttggggtt agggttagtt
     3061 taagttgaca tgtacttgca aagtttctta taatcagtta aatgtctgtt gaagtagcag
     3121 tatcaccaga tattaaactg acatacagtc tactaatact caaatggatc ataaaaatca
     3181 agggttacca ccagtttata tgaaatgtct ccagaatatg attaagctta ccactgggag
     3241 caaaggtgtt ctgaaattaa cacaatacat taacaacaca cattgttgtc atgggctatg
     3301 agcgtctgtg tagcagatca ggcccagccc tgtgtttatt ctcgtgcata gtgaagtccc
     3361 cacaatttct gaggagaaaa aaggaaaagg tccattggaa tgcccagagt aaggaggcaa
     3421 gaggccgccc tggtgctgcc cggattggag tgttttctcc cagtcatccc tataaggctt
     3481 tagtggagtg aggctaccga tccgagtgca gcccaggctc cttgtttggg atgttagagg
     3541 gtgtggtgag cagagatgga aactacataa gtacaccaca aaccagtgag gtgtacacca
     3601 agtttgtaca aaaaagcagg ctccaccatg gtgagcaagg gcgaggagga caacatggcc
     3661 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     3721 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     3781 aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     3841 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     3901 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     3961 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     4021 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     4081 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     4141 ctgaaggacg gcggccacta cgacgccgag gtcaagacca cctacaaggc caagaagccc
     4201 gtgcagctgc ccggcgccta caacgtcaac atcaagctgg acatcacctc ccacaacgag
     4261 gactacacca tcgtggaaca gtacgagcgc gccgagggcc gccactccac cggcggcatg
     4321 gacgagctgt acaagtaaga cccagctttc ttgtacaaag tgggggatcc agacatgata
     4381 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     4441 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     4501 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     4561 tccaacttat tatacatagt tgataattca ctggccgtcg ttttacggta ccatcgatga
     4621 tgatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     4681 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     4741 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     4801 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga
     4861 tcctctagat cagatctgcg aagatacggc cacgggtgct cttgatcctg tggctgattt
     4921 tggactgtgc tgctcgcagc tgctgatgaa tcacatactt cctccatttt cttccactga
     4981 ttgactgtta taatttccct aatttccagg tcaaggtgct gtgcattgtg gtaatagatg
     5041 tgacatgacg tcacttccaa aggaccaatg aacatgtctg accaatttca tataatgtga
     5101 aaacgatttt cataggcaga ataaataaca tttaaattaa actgggcatc agcgcaattc
     5161 aattggtttg gtaatagcaa gggaaaatag aatgaagtga tctccaaaaa ataagtactt
     5221 tttgactgta aataaaattg taaggagtaa aaagtacttt tttttctaaa aaaatgtaat
     5281 taagtaaaag taaaagtatt gatttttaat tgtactcaag taaagtaaaa atccccaaaa
     5341 ataatactta agtacagtaa tcaagtaaaa ttactcaagt actttacacc tctggttctt
     5401 gaccccctac cttcagcaag cccagcagat ccactagttc tagagcggcc gccaccgcgg
     5461 tggagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
     5521 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5581 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5641 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5701 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5761 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5821 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5881 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5941 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6001 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6061 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6121 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6181 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6241 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6301 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6361 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6421 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6481 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6541 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6601 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6661 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6721 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6781 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6841 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6901 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6961 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7021 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7081 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7141 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7201 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7261 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7321 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7381 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7441 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7501 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7561 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7621 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtg
//
