LOCUS       sequence-487082-        8890 bp    DNA     circular SYN 24-SEP-2025
DEFINITION  Gateway destination vector with the murine Mef2c anterior heart 
            field promoter in the backbone. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8890)
  AUTHORS   Gillespie W, Zhang Y, Ruiz OE, Cerda J, Ortiz-Guzman J, Turner WD, 
            Largoza G, Sherman M, Mosser LE, Fujimoto E, Chien CB, Kwan KM, 
            Arenkiel BR, Devine WP, Wythe JD
  TITLE     Multisite Assembly of Gateway Induced Clones (MAGIC): a flexible 
            cloning toolbox with diverse applications in vertebrate model 
            systems.
  JOURNAL   bioRxiv [Preprint]. 2024 Jul 13:2024.07.13.603267. doi: 
            10.1101/2024.07.13.603267.
  PUBMED    39026881
REFERENCE   2  (bases 1 to 8890)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2024 Jul 13:2024.07.13.603267. doi: 
            10.1101/2024.07.13.603267."
COMMENT     Sequence Label: pMef2c-AHF-DEST (JDW 476)
FEATURES             Location/Qualifiers
     source          1..8890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4057..4076
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    4116..4240
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(4265..4270,4271..4288,4289..4295)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             4349..5008
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4415..4434)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             5350..5655
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     5534..5553
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(5696..5820)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5912..5931)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5977..6032
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6031..6050)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6393..6409)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6393..6409)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6406..6428)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6417..6433
                     /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(6441..6447,6448..6465,6466..6471))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6486..6507
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6566..6761
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6612..6747
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6767..6901
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6804..6823)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6858..6877
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6969..6986)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7140..7728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7220..7239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7899..8690,8691..8759))
                     /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     8522..8541
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8760..8864)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tatcgagagg aaagccacca gtttctttgt cttgaagtat
       61 ttatttaaaa tgttggcatg tgtgacagtg atacagttaa atggccatgc tcttctttat
      121 gatatgtgca gtgctgtggc atggtggttg ctcagagctc ctaaaacctt ccaagggcca
      181 cctttgcaag tgcctactac aattaagctg attcctaggt ggccatgtac atcatcatag
      241 gacaaactac aaagaggtta aaatatcttt tacattatgg aatttgaatc ttaccatgtt
      301 tctcggattt ttatttttct aaattttcat gttcaattat atttatgatg aaagtattct
      361 ggctgaagaa caaatactat acacttgaat aatcttgagg ggtagattaa tctatatcct
      421 ttcttcatct cagggtaggt tgctgtggca accgccccag ctcactttaa aataattttt
      481 taaagctcac ttcaagagct acacccagcg tgggccaggc ttaaaagcag gcagacccag
      541 tgcatagggc tttcgtagac tgctgcttta atgtagggag gcagaagaaa gggagctgaa
      601 gtaagatggc tcaaaacaga aattggaatg ttagctttga gctgaaacaa tcaatcacca
      661 tgctctctgt aacttcaggt gtctaaatgc attgtgaaat aaatccttgg atgagataac
      721 taatgtttat ttatatttaa aattaagcac ttataataat taatgaaagg aatatttgta
      781 attacacaca ttcagagaga gaaggggagg gggggagaag gagagagagg aagtgagtga
      841 gtgattgaga gagagagaga gagagagagg gagggaggga gagggaggta aggagaaagg
      901 tggtttaatg gcaggtcaat tctagccatc acttcatctt ctttattaaa ggaaagtttc
      961 cttttattaa atccttagct tcactcccca ctgatataaa agatataata attatgatta
     1021 ttttcaagca tagaacaaaa ttggagagct catgcaaacc aggaaaccga ggcctagaaa
     1081 ggatttatct attaggaaga cacttttagt gttttcaatg taaactatca aaccataagt
     1141 aaaaaagtta gtagtgctct gccacagagt cacatctaca accacaaatc ccaaactgga
     1201 tagagcattc taagtctctc aacatctaga acaaaatact tcctacacaa gaaaaatgaa
     1261 taccattagg tagcaaggat caccagttta ggtaaataaa agtaaagaga taatgtgtgt
     1321 ctggcattac ttaaagacaa atataaaaca cactctggat taacaatctc atggattact
     1381 actaaattat ttacttattt atttacttat atatatattt atttattttc cacttcagct
     1441 acaaggtttg tgaggaaaac taacactaat cttaactttg ctcacacatc attaagaagc
     1501 taaatataca aagggggaaa atcttatctt ttcaaaggcc agctgtaata cattccagac
     1561 ccaaagtttc aaatcatcct cacctggact ggacatactt ttaaatgttt acaatattgt
     1621 tgataaatac gggtttgtta caatacttat tactaaacaa ttctttgaca cctggtgggg
     1681 aaaaataaaa gtttcctttt gaaggaaaat aaatcaaaac tgaagtgtgc tgtggtaagt
     1741 agaaactttg tctataatta aggagaagag catgattgaa ttcatgtatt ggctatatca
     1801 actctgtcat gtaacatagg ccattatagt tcattatact aatttaaagc actatgagaa
     1861 taagttctct tggtccttct tatagtttgc tctgtgagga tctttgtaaa ccttaaaaat
     1921 aatctaaatt tttatttata tcaccatatc taggtatgcc ttagcatgca gcatgtctaa
     1981 gctcagagct tgtacctcta aacatgccgt tctgtttata taaactcatg gtagtcacac
     2041 ttgctatgtg aaagaggagc aaatacaata tagcatagat accattttca aaagcagaac
     2101 aaacaaaaaa cccaccatgt cacaaagctt gtgctctgtg acagcttgtc atgctttcct
     2161 gtaataaaat cgcagtacta attaatagtc tcagtggtaa ggagccagag gtttgaagat
     2221 tgttcccatg tagtttaact tgcctgtaaa atcaactttc tgctttcaaa tgaagccttc
     2281 cttcggaggg tgcttaaatg ttgagtccac tgctctgata gaagctacta acttcaaaca
     2341 gcacaatgag gcaggaggct tttttgatgc acaatgcccc tctgcattaa aatagtactc
     2401 tgcaggccta ctggagagct tcccggggaa ttcagccagt tactgggtta aacatatact
     2461 ttcgaaaata tttttaatta gctagatatt tctatgggct actattgtct actcttgtat
     2521 cttctacatt atagtactct gtgaactttc tcttcctcta atcatagtat taaaaattaa
     2581 atgttcatat tttacacaag ctacagtact cctggcatgc aatctttcac tattttgaga
     2641 atgcataggg ttgacttgta tatagttgat agatttaatc tttagctttt taggtgtggt
     2701 cgggatatct agtacctttt ttttttctaa gtatcaattc tttgttaacc tatattttta
     2761 ctatggaaaa ttattcgaca gcttaaacct taacctaaga gttcttatca gtgtctgctc
     2821 ctgcttcctg tgcagctgaa actcttcagg ataagtgaag tcacccgcta tctatcggtc
     2881 aggggagcct aatgcatttg ggaagttgtt tttggtttac ttgctaatga cctggataaa
     2941 gtccagtaag gtgtggaggg aaggaggggg cattctccga ggaagctgtc ctctcagtaa
     3001 atttgtccag attgattcga aagttaatgg cccggggata aagtcgagat ttcaaaaact
     3061 tgtaattaat ttattactaa cattggagga tccattcggc tcactttcta gcctagccct
     3121 cctgactctc aaggccaaaa gctttataca ttctgggcgg gcaggaagcg cacgctggga
     3181 tcgaggcttg cagacatcct ggatcgtggc ttttacgcgt gagtaacgga ggcgcgcgcg
     3241 gaccggagca aggatgaagt ggctactgag aatctaaagg gggcgggggg ggggggggga
     3301 gtcactagag cagctcttag ggtcgcgctc ctattggtca gaaggaagat ctcgacttca
     3361 gaaggacttt cctgctagct ggcttgcagc ctccccgggg gagcgagcgc gcaggagact
     3421 agcgtcctct gtagcggcgg ggagagaggc agggagccgc cgctccccgt gttgtcttcc
     3481 ctttcatgca ataaagaatt gtcattaggt ttgcgtatgg catgtgccat taccccaccg
     3541 taaaactaaa attagcaaaa tgtcaggaat ggaaagatcg attcaccaag atgcaattat
     3601 catttaaaag tgcttgattg aggtactgat gttcagtgat ttattctgca taccatatac
     3661 ataattaaag tagtgtagtg gagtaattta tcaatctagt tgagactgga ggggtgagga
     3721 gggagctgca gtatgtttgt tttattaaaa tgttgcgagg ttctagtccc gccccccttt
     3781 tgcaagagtg aaactgatga tttccccagc tcgggaggag aggcaacggt ttgggctggt
     3841 tgggatagag aaacggaaag gaaaaggcag cgccaggcaa aggcaagaat aaaatgaatt
     3901 gggggacggg ggtagcaccg gagcgtacgt gctgcttaga accccagccc cgtgtgctcc
     3961 gtcttgcttg ccgcccccgc gtcccctccc agtcgaggtc gactagagcc tctgctaacc
     4021 atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt
     4081 ctcatcattt tggcaaagaa ttcctcgagg atatcacaag tttgtacaaa aaagctgaac
     4141 gagaaacgta aaatgatata aatatcaata tattaaatta gattttgcat aaaaaacaga
     4201 ctacataata ctgtaaaaca caacatatcc agtcactatg gcggccgcat taggcacccc
     4261 aggctttaca ctttatgctt ccggctcgta taatgtgtgg attttgagtt aggatccgtc
     4321 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
     4381 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
     4441 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
     4501 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
     4561 ggaattccgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
     4621 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
     4681 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
     4741 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt
     4801 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
     4861 caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
     4921 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
     4981 ctgcgatgag tggcagggcg gggcgtaaac gcgtggatcc ggcttactaa aagccagata
     5041 acagtatgcg tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac
     5101 ccgaagtatg tcaaaaagag gtatgctatg aagcagcgta ttacagtgac agttgacagc
     5161 gacagctatc agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa
     5221 ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag
     5281 ggatggctga ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg
     5341 gctggtgaaa tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt
     5401 gtggatgtac agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc
     5461 agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg
     5521 gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg
     5581 gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg
     5641 ttctggggaa tataaatgtc aggctccctt atacacagcc agtctgcagg tcgaccatag
     5701 tgactggata tgttgtgttt tacagtatta tgtagtctgt tttttatgca aaatctaatt
     5761 taatatattg atatttatat cattttacgt ttctcgttca gctttcttgt acaaagtggt
     5821 gatatcgcgg ccgcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg
     5881 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg
     5941 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     6001 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     6061 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     6121 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     6181 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     6241 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     6301 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc
     6361 tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6421 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6481 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6541 ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc
     6601 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     6661 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     6721 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc
     6781 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6841 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     6901 ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6961 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     7021 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     7081 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     7141 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     7201 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     7261 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     7321 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     7381 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7441 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7501 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7561 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7621 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7681 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7741 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7801 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7861 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7921 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7981 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     8041 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     8101 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     8161 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     8221 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     8281 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     8341 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     8401 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8461 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     8521 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8581 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8641 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8701 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8761 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8821 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8881 gtgccacctg
//
