LOCUS       sequence-248971-        5980 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 5980)
  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 5980)
  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
FEATURES             Location/Qualifiers
     source          1..5980
                     /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    2032..2166
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2056..2075)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2110..2129
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2279..2299)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2279..2297)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2318..2334)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2318..2334)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2331..2353)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2342..2358
                     /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(2366..2372,2373..2390,2391..2396))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2411..2432
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2549..2566)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2720..3308)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2800..2819)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3479..4270,4271..4339))
                     /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     4102..4121
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4340..4444)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4470..4925)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4607..4628)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4819..4838
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4996..5980
                     /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 cctctagaac tatagtgagt cgtattacgt agatccagac
     2041 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     2101 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     2161 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     2221 gttttttaat tcgcggccgc ggcgccaatg cattgggccc ggtacccagc ttttgttccc
     2281 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     2341 attgttatcc gctcacaatt ccacacaaca tacgagccgg gagcataaag tgtaaagcct
     2401 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     2461 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2521 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2581 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2641 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2701 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2761 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2821 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2881 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2941 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3001 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3061 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3121 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3181 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3241 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3301 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3361 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3421 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3481 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3541 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3601 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3661 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3721 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3781 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3841 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3901 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3961 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4021 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4081 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4141 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4201 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4261 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4321 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4381 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4441 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     4501 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     4561 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     4621 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     4681 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     4741 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     4801 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     4861 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4921 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     4981 ttcgctatta cgccagtcga ccgccaattc aatatggcgt atatggactc atgccaattc
     5041 aatatggtgg atctggacct gtgccaattc aatatggcgt atatggactc gtgccaattc
     5101 aatatggtgg atctggaccc cagccaattc aatatggcgg acttggcacc atgccaattc
     5161 aatatggcgg acctggcact gtgccaactg gggaggggtc tacttggcac ggtgccaagt
     5221 ttgaggaggg gtcttggccc tgtgccaagt ccgccatatt gaattggcat ggtgccaata
     5281 atggcggcca tattggctat atgccaggat caatatatag gcaatatcca atatggccct
     5341 atgccaatat ggctattggc caggttcaat actatgtatt ggccctatgc catatagtat
     5401 tccatatatg ggttttccta ttgacgtaga tagcccctcc caatgggcgg tcccatatac
     5461 catatatggg gcttcctaat accgcccata gccactcccc cattgacgtc aatggtctct
     5521 atatatggtc tttcctattg acgtcatatg ggcggtccta ttgacgtata tggcgcctcc
     5581 cccattgacg tcaattacgg taaatggccc gcctggctca atgcccattg acgtcaatag
     5641 gaccacccac cattgacgtc aatgggatgg ctcattgccc attcatatcc gttctcacgc
     5701 cccctattga cgtcaatgac ggtaaatggc ccacttggca gtacatcaat atctattaat
     5761 agtaacttgg caagtacatt actattggaa gtacgccagg gtacattggc agtactccca
     5821 ttgacgtcaa tggcggtaaa tggcccgcga tggctgccaa gtacatcccc attgacgtca
     5881 atggggaggg gcaatgacgc aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc
     5941 ctaatgggag gtctatataa gcaatgctcg tttagggaac
//
