LOCUS       sequence-248968-        6790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mRNA synthesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6790)
  AUTHORS   Horb ME, Thomsen GH
  TITLE     Tbx5 is essential for heart development.
  JOURNAL   Development. 1999 Apr;126(8):1739-51.
  PUBMED    10079235
REFERENCE   2  (bases 1 to 6790)
  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: 
            "Development."; date: "1999-04"; volume: "126(8)"; pages: "1739-51"
COMMENT     Sequence Label: pCS2-XTbx5-EnR-GR
FEATURES             Location/Qualifiers
     source          1..6790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    2842..2976
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2866..2885)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2920..2939
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3089..3109)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3089..3107)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3128..3144)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3128..3144)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3141..3163)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3152..3168
                     /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(3176..3182,3183..3200,3201..3206))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3221..3242
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3359..3376)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3530..4118)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3610..3629)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4289..5080,5081..5149))
                     /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     4912..4931
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5150..5254)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5280..5735)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5417..5438)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5629..5648
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5806..6790
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattag cttttccaag agaagaaaag
      121 tttttatggt gtggctcagg gacctgtacc tgagagccca ggagcccagg ataccttctc
      181 cctttgcaca gtgtttgcca ttggcaatcc cttgccagtg ccactgcctg aagtgcaggc
      241 tcccactgga atatggcgga cacagaggag gcttatggga tgcctgatac ccctgtggaa
      301 gcagagccca aagaactaca gtgtgagcct aaacaggaca atcaattggg ggcaagcagc
      361 aagaccccca cttctcctcc ggctgcattt acccagcagg gaatggaagg cattaaggtg
      421 tttttgcatg agagagaact atggctgaaa tttcatgaag ttggcacaga aatgattatt
      481 actaaggctg gaaggcgcat gttccctagt tacaaagtga aggttactgg cctgaaccca
      541 aaaaccaagt acatcctcct catggatatc gttcctgcag atgaccacag atacaagttt
      601 gctgataaca aatggtcagt cacaggcaag gctgaacccg ctatgccagg acgtttgtat
      661 gttcacccag attcaccagc cacgggcgca cattggatga ggcagctagt ctccttccag
      721 aagctgaagc tcacaaacaa ccatctggat ccatttggac atatcatcct gaattcaatg
      781 cacaagtacc agccacggct ccacattgtc aaggcagatg agaacaatgg ctttgggtcc
      841 aagaatacag ctttctgcac tcatgtattt tctgagactg attttattgc tgtcacctca
      901 tatcagaacc acaagatcac acagctaaag atagagaaca atccctttgc caaaggattt
      961 cggggaagtg atgacatgga gctacaccgg atgtccagga tgcagagcaa agagtatcca
     1021 gtagtgccaa gaagcacagt ccgtcagaaa gtctcctcta atcacagccc attctctcaa
     1081 gaaacacgaa ttcaaggtat ggccctggag gatcgctgca gcccacagtc agcgcccagc
     1141 cccattaccc tacaaatgca gcatcttcac caccagcaac agcagcagca gcaacagcag
     1201 cagcaaatgc agcacctcca ccagctgcag caactgcagc agttgcacca acagcaactg
     1261 gccgccggtg tcttccacca tccggcaatg gccttcgatg ccgctgcagc cgccgctgct
     1321 gcagctgctg ctgcggccgc ccacgctcat gctgctgcac tgcagcagcg cctcagtggc
     1381 agtggatcgc ccgcatcctg ctccacgccc gcctcgtcca cgccgctgac catcaaggag
     1441 gaggaaagcg actccgtgat cggtgacatg agtttccaca atcagacgca caccaccaac
     1501 gaggaggagg aggcggagga ggatgacgac attgatgtgg atgtggatga tacgtcggcg
     1561 ggcggacgcc tgccaccacc cgcccaccag cagcagtcga cggccaagcc ctcgctggcc
     1621 ttttccatct ccaacatcct gagcgatcgt ttcggagatg tccagaagcc tggcaagtcg
     1681 attgagaacc aggccagcat attccgcccc ttcgaggcga atcgctccca gactgccacg
     1741 ccctccgcct ttacaagagt ggatctgctg gagtttagcc ggcaacagca agctgccgcc
     1801 gcagccgcta ctgcggccat gatgctggaa cgggccaact tccttaactg cttcaatccg
     1861 gctgcctatc ccaggataca cgaggaaatc gtgcagagtc gcctgcgcag gagtgcagcc
     1921 aatgccgtca tcccgccgcc catgagctcc aagatgagcg atgccaatcc agagaaatct
     1981 gctctgggat ccctctcgag cccctctgaa aatcctggta acaaaacaat agttcctgca
     2041 acgttaccac aactcacccc taccctggtg tcactgttgg aggttattga acctgaagtg
     2101 ttatatgcag gatatgatag ctctgttcca gactcaactt ggaggatcat gactacgctc
     2161 aacatgttag gagggcggca agtgattgca gcagtgaaat gggcaaaggc aataccaggt
     2221 ttcaggaact tacacctgga tgaccaaatg accctactgc agtactcctg gatgtttctt
     2281 atggcatttg ctctggggtg gagatcatat agacaatcaa gtgcaaacct gctgtgtttt
     2341 gctcctgatc tgattattaa tgagcagaga atgactctac cctgcatgta cgaccaatgt
     2401 aaacacatgc tgtatgtttc ctctgagtta cacaggcttc aggtatctta tgaagagtat
     2461 ctctgtatga aaaccttact gcttctctct tcagttccta aggacggtct gaagagccaa
     2521 gagctatttg atgaaattag aatgacctac atcaaagagc taggaaaagc cattgtcaag
     2581 agggaaggaa actccagcca gaactggcag cggttttatc aactgacaaa actcttggat
     2641 tctatgcatg aagtggttga aaatctcctt aactattgct tccaaacatt tttggataag
     2701 accatgagta ttgaattccc cgagatgtta gctgaaatca tcaccaatca gataccaaaa
     2761 tattcaaatg gaaatatcaa aaaacttctg tttcatcaaa agtgctcgag cctctagaac
     2821 tatagtgagt cgtattacgt agatccagac atgataagat acattgatga gtttggacaa
     2881 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     2941 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     3001 atgtttcagg ttcaggggga ggtgtgggag gttttttaat tcgcggccgc ggcgccaatg
     3061 cattgggccc ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     3121 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     3181 tacgagccgg gagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     3241 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3301 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3361 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3421 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3481 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3541 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3601 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3661 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3721 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3781 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3841 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3901 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3961 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4021 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4081 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4141 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4201 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4261 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4321 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4381 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4441 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4501 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4561 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4621 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4681 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4741 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4801 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4861 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4921 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4981 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5041 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5101 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5161 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5221 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     5281 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     5341 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     5401 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     5461 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     5521 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     5581 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     5641 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     5701 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
     5761 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagtcga ccgccaattc
     5821 aatatggcgt atatggactc atgccaattc aatatggtgg atctggacct gtgccaattc
     5881 aatatggcgt atatggactc gtgccaattc aatatggtgg atctggaccc cagccaattc
     5941 aatatggcgg acttggcacc atgccaattc aatatggcgg acctggcact gtgccaactg
     6001 gggaggggtc tacttggcac ggtgccaagt ttgaggaggg gtcttggccc tgtgccaagt
     6061 ccgccatatt gaattggcat ggtgccaata atggcggcca tattggctat atgccaggat
     6121 caatatatag gcaatatcca atatggccct atgccaatat ggctattggc caggttcaat
     6181 actatgtatt ggccctatgc catatagtat tccatatatg ggttttccta ttgacgtaga
     6241 tagcccctcc caatgggcgg tcccatatac catatatggg gcttcctaat accgcccata
     6301 gccactcccc cattgacgtc aatggtctct atatatggtc tttcctattg acgtcatatg
     6361 ggcggtccta ttgacgtata tggcgcctcc cccattgacg tcaattacgg taaatggccc
     6421 gcctggctca atgcccattg acgtcaatag gaccacccac cattgacgtc aatgggatgg
     6481 ctcattgccc attcatatcc gttctcacgc cccctattga cgtcaatgac ggtaaatggc
     6541 ccacttggca gtacatcaat atctattaat agtaacttgg caagtacatt actattggaa
     6601 gtacgccagg gtacattggc agtactccca ttgacgtcaa tggcggtaaa tggcccgcga
     6661 tggctgccaa gtacatcccc attgacgtca atggggaggg gcaatgacgc aaatgggcgt
     6721 tccattgacg taaatgggcg gtaggcgtgc ctaatgggag gtctatataa gcaatgctcg
     6781 tttagggaac
//
