LOCUS       sequence-266110-        6618 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transgenesis vector consisting of a 1.6-kb fragment of MyHC1 
            promoter region upstream of the start codon and an mCherry reporter 
            gene, flanked by inverted binding sites of meganuclease I-SceI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6618)
  AUTHORS   Renfer E, Amon-Hassenzahl A, Steinmetz PR, Technau U
  TITLE     A muscle-specific transgenic reporter line of the sea anemone, 
            Nematostella vectensis.
  JOURNAL   Proc Natl Acad Sci U S A. 2010 Jan 5;107(1):104-8. doi: 
            10.1073/pnas.0909148107. Epub 2009 Dec 14.
  PUBMED    20018670
REFERENCE   2  (bases 1 to 6618)
  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"; doi: 
            "10.1073/pnas.0909148107"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2010-01-5- 5"; volume: "107"; issue: "1"; pages: "104-8"
COMMENT     Sequence Label: pNvT-MHC::mCh
FEATURES             Location/Qualifiers
     source          1..6618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    10..31
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(46..51,52..69,70..76)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    84..100
                     /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     89..111
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     108..124
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     108..124
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        142..160
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     142..159
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1900..2610
                     /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(2045..2063)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2313..2332
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2732..2853
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2769..2788)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2823..2842
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2954..2973)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2955..2973)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2980..2997)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2980..2996)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2989..3011)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3137..3565
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3224..3243)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3434..3455
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3909..4703
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3963..3982)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4573..4592
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4721..4789,4790..5581)
                     /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     complement(4939..4958)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5752..6340
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6241..6260
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6494..6511
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
       61 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
      121 tgaccatgat tacgccaagc tatttaggtg acactataga atactcaagc tatgcatcaa
      181 gcttggtacc gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcgcccttg
      241 atagggataa cagggtaatt taattaaccg gatgggagca aagaaaaact tgtctgcaac
      301 acctaagtag aaattccctg ctagcagagg cctcttgtct cgtgttttcg atgggctaac
      361 gctgggctaa cgccagccca gcgaaataca gagataatag gcctctagca gggaaaagct
      421 gtaacactca tctcctaact tttatttcaa atcattttct ataaaaaaat gcatagaaat
      481 ttctacctta ctcacattaa cgagatcaat cgctggaaca tatggaaatg ttagcttaca
      541 tcgacacctt tttttcattc cacctaaaaa tatcagaaaa attcttatca gcacagtagc
      601 actgtgaata accccaaatt tagacaccat agggaacgcg atttattcac accatggtac
      661 caaacccgca agatgtactt gccagtgaat tgaagtaaaa attaaacaaa acgcaacgtg
      721 atttctaaaa atcctcgtga tcaaatttaa aaggacggtt ttatcattga agagtgctat
      781 tggtattgcg ttactgcgac catttctcta gtcacgttac gataccgtga cacgatacat
      841 cggtaaataa aactgtcctt gacaaaataa ctctccctca ggttcatatc gcgctgaaca
      901 aatcctagaa aagggcgacg attacataga tccgcgctgt taaatccaaa tagacacgcg
      961 cctcgtagaa gcctgacggg agcccagctc ggtcaccgct actcgtaaaa ccgtgtcagg
     1021 gttgaagtaa ttatacgtgt cattctgatg cgatgtttat tatgatacgg ttcggtgacg
     1081 gccatcagac cactctaatt ctgtcgctag catcttttga ttctctctta aagcacaata
     1141 aactgcgcta agggtcggtt tgcatggccc acagaagcat tcagccgcga gacaagcgag
     1201 taatttcctg atgggaagcg ccggtcgtga aacacacgtg ccttgctgct acaaatcaag
     1261 ataacgctct aaatgcggta caatacactc cacttgtact gttgggcaat ttattgctat
     1321 tggaagtaat ttatttggac tgaataaaat aattttgcga aaagacggga cagggtcgta
     1381 ttctcgggta tgtttgaatg gatttgattt gggcgataaa aagcgttatg ggggcatttt
     1441 atttgtgcaa ttttcagaaa ttgttcacat gctgatagtg tacagcctag cgttaaatcg
     1501 gccgatgaga atcggatcag aactgggtac gctgattggg cagtgggccc aaggtgtggt
     1561 aaaacagtga taatagtgat tgtttggtcg acctgatgcg gcaaaccgtg acaaaataaa
     1621 ttccgtcgat ctcgggacaa tgcgctaagt caaacgagag ccaatcagag agcgggatct
     1681 tgaccgaaat acctagaggt cgcgtctgtt atgcgaaccg ggagtggctt agggggttta
     1741 tagagcgggc tggatcgtac tcaacactag ctcagagtgg caagcacctt ctgcggacac
     1801 actgaagctg ctagcacgct aagcgacaac actcggtatt cttctccaaa tatatttgta
     1861 tttcacttgt gtttagaata actgtttcaa gggcgcgcca tggtgagcaa gggcgaggag
     1921 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     1981 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     2041 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     2101 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     2161 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     2221 ggcgtggtga ccgtgaccca ggactcctcc ctccaggacg gcgagttcat ctacaaggtg
     2281 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     2341 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     2401 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     2461 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     2521 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     2581 accggcggca tggacgagct gtacaagtaa cctgcaggga ctctagatca taatcagcca
     2641 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     2701 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     2761 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2821 ttgtggtttg tccaaactca tcaatgtatc ttaaagggcc ggccattacc ctgttatccc
     2881 tatcaagggc gaattctgca gatatccatc acactggcgg ccgctcgagc atgcatctag
     2941 agggcccaat tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg
     3001 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     3061 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     3121 cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3181 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3241 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3301 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3361 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3421 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3481 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3541 atttaacaaa aatttaacgc gaattttaac aaaattcagg gcgcaagggc tgctaaagga
     3601 agcggaacac gtagaaagcc agtccgcaga aacggtgctg accccggatg aatgtcagct
     3661 actgggctat ctggacaagg gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg
     3721 ggcttacatg gcgatagcta gactgggcgg ttttatggac agcaagcgaa ccggaattgc
     3781 cagctggggc gccctctggt aaggttggga agccctgcaa agtaaactgg atggctttct
     3841 tgccgccaag gatctgatgg cgcaggggat caagatctga tcaagagaca ggatgaggat
     3901 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3961 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4021 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4081 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4141 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4201 cggggcagga tctcctgtca tcccaccttg ctcctgccga gaaagtatcc atcatggctg
     4261 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4321 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4381 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4441 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4501 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4561 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4621 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4681 gccttcttga cgagttcttc tgaattgaaa aaggaagagt atgagtattc aacatttccg
     4741 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4801 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4861 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4921 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4981 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5041 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5101 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5161 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5221 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5281 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5341 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5401 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5461 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5521 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5581 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5641 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5701 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5761 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5821 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5881 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5941 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6001 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6061 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6121 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6181 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6241 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6301 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6361 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6421 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     6481 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     6541 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     6601 gaaagcgggc agtgagcg
//
