LOCUS       Exported                8883 bp ds-DNA     circular SYN 02-MAR-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8883)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jul 15, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8883
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        19..3988
                     /label=Mef2C AHF Promoter
     protein_bind    complement(2030..2037)
                     /label=Tbx5 Binding Site (ECR Browser)
     primer_bind     3710..3729
                     /label=Mef2c-AHF-Pro-Fwd
     primer_bind     3916..3935
                     /label=WPD-Seq2
     intron          3989..4107
                     /label=Rabbit Globin Intron
     intron          4045..4097
                     /label=rabbit B-globin intron
     gene            4108..5818
                     /label=ccdB RFA
     protein_bind    4111..4235
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        4260..4290
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             4344..5003
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             5345..5650
                     /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"
     protein_bind    complement(5691..5815)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_site      5822..6364
                     /label=Rabbit Globin pA
     polyA_site      5830..6373
                     /label=Rabbit globin polyA
     polyA_signal    5972..6027
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6382..6402)
                     /label=M13-rev
     misc_binding    complement(6408..6430)
                     /label=LacO
     promoter        complement(6434..6464)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6479..6500
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6559..6754
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6605..6740
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6760..6894
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_site      6760..6890
                     /label=SV40 early pA
     rep_origin      complement(7115..7797)
                     /direction=LEFT
                     /label=ColE1 origin
     CDS             complement(7892..8752)
                     /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     8684..8703
                     /label=WPD-Seq1
     promoter        complement(8753..8857)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgacg agaggaaagc caccagtttc tttgtcttga agtatttatt
       61 taaaatgttg gcatgtgtga cagtgataca gttaaatggc catgctcttc tttatgatat
      121 gtgcagtgct gtggcatggt ggttgctcag agctcctaaa accttccaag ggccaccttt
      181 gcaagtgcct actacaatta agctgattcc taggtggcca tgtacatcat cataggacaa
      241 actacaaaga ggttaaaata tcttttacat tgtggaattt gaatcttacc atgtttctcg
      301 gatttttatt tttctaaatt ttcatgttca attatattta tgatgaaagt attctggctg
      361 aagaacaaat actatacact tgaataatct tgaggggtag attaatctat atcctttctt
      421 catctcaggg taggttgctg tggcaaccgc cccagctcac tttaaaataa ttttttaaag
      481 ctcacttcaa gagctacacc cagcgtgggc caggcttaaa agcaggcaga cccagtgcat
      541 agggctttcg tagactgctg ctttaatgta gggaggcaga agaaagggag ctgaagtaag
      601 atggctcaaa acagaaattg gaatgttagc tttgagctga aacaatcaat caccatgctc
      661 tctgtaactt caggtgtcta aatgcattgt gaaataaatc cttggatgag ataactaatg
      721 tttatttata tttaaaatta agcacttata ataattaatg aaaggaatat ttgtaattac
      781 acacattcag agagagaagg ggagaggggg agaaggagag agaggaagtg agtgagtgat
      841 tgagagagag agagagagag agagggaggg agggagaggg aggtaaggag aaaggtggtt
      901 taatggcagg tcaattctag ccatcacttc atcttcttta ttaaaggaaa gtttcctttt
      961 attaaatcct tagcttcact ccccactgat ataaaagata taataattat gattattttc
     1021 aagcatagaa caaaattgga gagctcatgc aaaccaggaa accgaggcct agaaaggatt
     1081 tatctattag gaagacactt ttagtgtttt caatgtaaac tatcaaacca taagtaaaaa
     1141 agttagtagt gctctgccac agagtcacat ctacaaccac aaatcccaaa ctggatagag
     1201 cattctaagt ctctcaacat ctagaacaaa atacttccta cacaagaaaa atgaatacca
     1261 ttaggtagca aggatcacca gtttaggtaa ataaaagtaa agagataatg tgtgtctggc
     1321 attacttaaa gacaaatata aaacacactc tggattaaca atctcatgga ttactactaa
     1381 attatttact tatttattta cttatatata tatttattta ttttccactt cagctacaag
     1441 gtttgtgagg aaaactaaca ctaatcttaa ctttgctcac acatcattaa gaagctaaat
     1501 atacaaaggg ggaaaatctt atcttttcaa aggccagctg taatacattc cagacccaaa
     1561 gtttcaaatc atcctcacct ggactggaca tacttttaaa tgtttacaat attgttgata
     1621 aatacgggtt tgttacaata cttattacta aacaattctt tgacacctgg tggggaaaaa
     1681 taaaagtttc cttttgaagg aaaataaatc aaaactgaag tgtgctgtgg taagtagaaa
     1741 ctttgtctat aattaaggag aagagcatga ttgaattcat gtattggcta tatcaactct
     1801 gtcatgtaac ataggccatt atagttcatt atactaattt aaagcactat gagaataagt
     1861 tctcttggtc cttcttatag tttgctctgt gaggatcttt gtaaacctta aaaataatct
     1921 aaatttttat ttatatcacc atatctaggt atgccttagc atgcagcatg tctaagctca
     1981 gagcttgtac ctctaaacat gccgttctgt ttatataaac tcatggtagt cacacttgct
     2041 atgtgaaaga ggagcaaata caatatagca tagataccat tttcaaaagc agaacaaaca
     2101 aaaaacccac catgtcacaa agcttgtgct ctgtgacagc ttgtcatgct ttcctgtaat
     2161 aaaatcgcag tactaattaa tagtctcagt ggtaaggagc cagaggtttg aagattgttc
     2221 ccatgtagtt taacttgcct gtaaaatcaa ctttctgctt tcaaatgaag ccttccttcg
     2281 gagggtgctt aaatgttgag tccactgctc tgatagaagc tactaacttc aaacagcaca
     2341 atgaggcagg aggctttttt gatgcacaat gcccctctgc attaaaatag tactctgcag
     2401 gcctactgga gagcttcccg ggaaattcag ccagttactg ggttaaacat atactttcga
     2461 aaatattttt aattagctag atatttctat gggctactat tgtctactct tgtatcttct
     2521 acattatagt actctgtgaa ctttctcttc ctctaatcat agtattaaaa attaaatgtt
     2581 catattttac acaagctaca gtactcctgg catgcaatct ttcactattt tgagaatgca
     2641 tagggttgac ttgtatatag ttgatagatt taatctttag ctttttaggt gtggtcggga
     2701 tatctagtac cttttttttt tctaagtatc aattctttgt taacctatat ttttactatg
     2761 gaaaattatt cgacagctta aaccttaacc taagagttct tatcagtgtc tgctcctgct
     2821 tcctgtgcag ctgaaactct tcaggataag tgaagtcacc cgctatctat cggtcagggg
     2881 agcctaatgc atttgggaag ttgtttttgg tttacttgct aatgacctgg ataaagtcca
     2941 gtaaggtgtg gagggaagga gggggcattc tccgaggaag ctgtcctctc agtaaatttg
     3001 tccagattga ttcgaaagtt aatggcccgg ggataaagtc gagatttcaa aaacttgtaa
     3061 ttaatttatt actaacattg gaggatcctt tcggctcact ttctagccta gccctcctga
     3121 ctctcaaggc caaaagcttt atacattctg ggcgggcagg aagcgcacgc tgggatcgag
     3181 gcttgcagac atcctggatc gtggctttta cgcgtgagta acggaggcgc gcgcggaccg
     3241 gagcaaggat gaagtggcta ctgagaatct aaagggggcg gggggggggg ggggagtcac
     3301 tagagcagct cttagggtcg cgctcctatt ggtcagaagg aagatctcga cttcagaagg
     3361 actttcctgc tagctggctt gcagcctccc cgggggagcg agcgcgcagg agactagcgt
     3421 cctctgtagc ggcggggaga gaggcaggga gccgccgctc cccgtgttgt cttccctttc
     3481 atgcaataaa gaattgtcat taggtttgcg tatggcatgt gccattaccc caccgtaaaa
     3541 ctaaaattag caaaatgtca ggaatggaaa gattgattca ccaagatgca attatcattt
     3601 aaaagtgctt gattgaggta ctgatgttca gtgatttatt ctgcatacca tatacataat
     3661 taaagtagtg tagtggagta atttatcaat ctagttgaga ctggaggggt gaggagggag
     3721 ctgcagtatg tttgttttat taaaatgttg cgaggttcta gtcccgcccc ccttttgcaa
     3781 gagtgaaact gatgatttcc ccagctcggg aggagaggca acggtttggg ctggttggga
     3841 tagagaaacg gaaaggaaaa ggcagcgcca ggcaaaggca agaataaaat gaattggggg
     3901 acgggggtag caccggagcg tacgtgctgc ttagaacccc agccccgtgt gctccgtctt
     3961 gcttgccgcc cccgcgtccc ctcccagtcg aggtcgacta gagcctctgc taaccatgtt
     4021 catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat
     4081 cattttggca aagaattcct cgaggatatc acaagtttgt acaaaaaagc tgaacgagaa
     4141 acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa acagactaca
     4201 taatactgta aaacacaaca tatccagtca ctatggcggc cgcattaggc accccaggct
     4261 ttacacttta tgcttccggc tcgtataatg tgtggatttt gagttaggat ccgtcgagat
     4321 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
     4381 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
     4441 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
     4501 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
     4561 tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
     4621 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
     4681 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
     4741 atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt
     4801 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
     4861 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
     4921 atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
     4981 atgagtggca gggcggggcg taaacgcgtg gatccggctt actaaaagcc agataacagt
     5041 atgcgtattt gcgcgctgat ttttgcggta taagaatata tactgatatg tatacccgaa
     5101 gtatgtcaaa aagaggtatg ctatgaagca gcgtattaca gtgacagttg acagcgacag
     5161 ctatcagttg ctcaaggcat atatgatgtc aatatctccg gtctggtaag cacaaccatg
     5221 cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg
     5281 gctgaggtcg cccggtttat tgaaatgaac ggctcttttg ctgacgagaa caggggctgg
     5341 tgaaatgcag tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga
     5401 tgtacagagt gatattattg acacgcccgg gcgacggatg gtgatccccc tggccagtgc
     5461 acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata tcggggatga
     5521 aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta tcggggaaga
     5581 agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg
     5641 gggaatataa atgtcaggct cccttataca cagccagtct gcaggtcgac catagtgact
     5701 ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata
     5761 tattgatatt tatatcattt tacgtttctc gttcagcttt cttgtacaaa gtggtgatat
     5821 cgcggccgca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat
     5881 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
     5941 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
     6001 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
     6061 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgccatatg ctggctgcca
     6121 tgaacaaagg tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc
     6181 cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt
     6241 atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct
     6301 cctctcctga ctactcccag tcatagctgt ccctcttctc ttatgaagat ccctcgacct
     6361 gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6421 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6481 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6541 tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     6601 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     6661 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     6721 aggaggcttt tttggaggcc taggcttttg caaaaagcta acttgtttat tgcagcttat
     6781 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     6841 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgctgc
     6901 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6961 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     7021 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     7081 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7141 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7201 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     7261 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     7321 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     7381 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     7441 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     7501 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     7561 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7621 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     7681 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7741 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7801 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7861 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7921 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7981 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     8041 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     8101 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     8161 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     8221 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     8281 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     8341 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     8401 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     8461 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     8521 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     8581 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     8641 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     8701 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8761 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8821 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8881 ctg
//
