LOCUS       sequence-249012-        7038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mRNA synthesis          in situ hyb probes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7038)
  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 7038)
  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
FEATURES             Location/Qualifiers
     source          1..7038
                     /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"
     primer_bind     complement(3040..3056)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3041..3057)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3090..3224
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3114..3133)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3168..3187
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3337..3357)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3337..3355)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3376..3392)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3376..3392)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3389..3411)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3400..3416
                     /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(3424..3430,3431..3448,3449..3454))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3469..3490
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3607..3624)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3778..4366)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3858..3877)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4537..5328,5329..5397))
                     /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     5160..5179
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5398..5502)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5528..5983)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5665..5686)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5877..5896
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6054..7038
                     /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 ttcgaatttt ttccaagaga agaaaagttt
      121 ttatggtgtg gctcagggac ctgtacctga gagcccagga gcccaggata ccttctccct
      181 ttgcacagtg tttgccattg gcaatccctt gccagtgcca ctgcctgaag tgcaggctcc
      241 cactggaata tggcggacac agaggaggct tatgggatgc ctgatacccc tgtggaagca
      301 gagcccaaag aactacagtg tgagcctaaa caggacaatc aattgggggc aagcagcaag
      361 acccccactt ctcctccggc tgcatttacc cagcagggaa tggaaggcat taaggtgttt
      421 ttgcatgaga gagaactatg gctgaaattt catgaagttg gcacagaaat gattattact
      481 aaggctggaa ggcgcatgtt ccctagttac aaagtgaagg ttactggcct gaacccaaaa
      541 accaagtaca tcctcctcat ggatatcgtt cctgcagatg accacagata caagtttgct
      601 gataacaaat ggtcagtcac aggcaaggct gaacccgcta tgccaggacg tttgtatgtt
      661 cacccagatt caccagccac gggcgcacat tggatgaggc agctagtctc cttccagaag
      721 ctgaagctca caaacaacca tctggatcca tttggacata tcatcctgaa ttcaatgcac
      781 aagtaccagc cacggctcca cattgtcaag gcagatgaga acaatggctt tgggtccaag
      841 aatacagctt tctgcactca tgtattttct gagactgatt ttattgctgt cacctcatat
      901 cagaaccaca agatcacaca gctaaagata gagaacaatc cctttgccaa aggatttcgg
      961 ggaagtgatg acatggagct acaccggatg tccaggatgc agagcaaaga gtatccagta
     1021 gtgccaagaa gcacagtccg tcagaaagtc tcctctaatc acagcccatt ctctcaagaa
     1081 acacgtaaca ttacaggatc ttctacccta aattcccaat atcagtgtga gaatggtgtt
     1141 agcagcacat ctcaggatct tctgccatct tccagtgcat atacttccct gccccatgag
     1201 tctggtacta tatatcactg cacaaaaaga aaagtaagtg aagagccagc tgagcattcc
     1261 tataagaaac cctacatgga cacttcacca agcgaagagg acccttttta ccgatctggg
     1321 tacccacaac cttcatcctc ttcctcatct accacctctt tccgcacaga atctgcccaa
     1381 cgacaggcct gcatgtatgc tagttcagca cctgccacag agcctgtacc atccatagag
     1441 gatatatctt gcaacagctg gtctagtgtg ccctcttaca gcagctgtac agtaggtgga
     1501 ggaatgcaac ccatggagcg cctcccatat cagcacttct ctgctcactt tacctctagt
     1561 tccctcatgc ctagactgag taatcatgca ggcacacagc caagtgattc tcactccatg
     1621 ttccaacatc agtcttctca tcaggccatt gtaaggcaat gcaatcctca atctgggtta
     1681 caacaatcat cagctcttca gcccactgaa ttcttgtacc cacacagtgt tccccggacc
     1741 atttcaccac atcaatacca ctctgtgcat ggggtaggta tggtcccaga ctggaatgaa
     1801 aacagctaac agggcagaaa gatgtggttt tggagtgcaa aaccgatatg ttgttcagag
     1861 agccattgaa tgtggacaga tgttggaagg atgcaaggat ttgtgtactg gaacctattt
     1921 cagtgatcca ctttcttcct aaactctcta gctttgtgat cctacataga aagaagaata
     1981 atacaaagtt ggagtgttcc taaaaggatt cacactcaaa aaaatagaga cctctcaaat
     2041 cctgtagctt tgcacttcat gcagtgggct aacactatat tgtggcatcc ttgtggaagg
     2101 acaaataaag caaaaaggag gggcaaaaag atgcacttaa tgtaagcacc tacacggctc
     2161 tggtgtttgc agtgtattac tcagtgtgct gtaggagaat ggaacagtct atgtaaaaac
     2221 aatgatgatg ctgtgtattc tgtcaaggag gacatggaaa gattgctctg gagaaaaagc
     2281 agagcttgta attagattcc tacacttcag caagtcttcg gagttgggtc accctctagg
     2341 aaagagacca atgttaagaa tatttaatgt tcatccgaaa tcaataacta aaaacatggg
     2401 aggaaaacat atgaataaat ccttgatctg tactgtagat tggtgatggt tgcagatacc
     2461 gccatttcag tatttgatgc atgacagctt atataattgg aatcttgtgg tcatttgtag
     2521 tattgctttt gtaaaaaatt ggagtaaaaa aattacaatt tctgttctgc aatagttcta
     2581 catagctatg ttgctattcc acccaatttc attccaattc aaagacattg gctactatct
     2641 attgtctacc aagatcgtca agaacataat gcaaatatgc atatcatgga agtatggttt
     2701 ctcttctgcc acagaaatga gtaatatcat gccaagttaa accttccttc ccacaattac
     2761 ctttttttgt aagaaaacct ccataagtta tttttaatta ttactttgtg acccaattgc
     2821 taaataacgt tgacctttgg ctacataatt gggtggtcaa cacttccaac agctgcagcc
     2881 atcctacaac gggcctgaac aatggatcta gcctgcacct cacctccaca taaaacagag
     2941 tgcttggttt gtcaaatgtt gtgtcattga ccttgttata taataaaaga taatatattt
     3001 ttgaaaaaaa aaaaaaacgg aattcgatat caagcttatc gataccgtcg acctcgagcc
     3061 tctagaacta tagtgagtcg tattacgtag atccagacat gataagatac attgatgagt
     3121 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3181 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     3241 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaattc gcggccgcgg
     3301 cgccaatgca ttgggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg
     3361 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3421 acacaacata cgagccggga gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3481 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3541 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3601 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3661 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3721 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3781 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3841 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3901 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3961 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4021 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4081 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4141 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4201 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4261 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4321 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4381 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4441 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4501 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4561 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4621 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4681 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4741 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4801 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4861 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4921 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4981 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5041 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5101 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5161 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5221 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5281 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5341 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5401 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5461 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5521 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     5581 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     5641 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     5701 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     5761 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     5821 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     5881 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     5941 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg
     6001 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagtcgacc
     6061 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
     6121 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
     6181 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
     6241 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
     6301 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
     6361 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
     6421 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
     6481 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
     6541 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
     6601 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
     6661 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
     6721 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
     6781 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
     6841 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
     6901 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
     6961 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
     7021 aatgctcgtt tagggaac
//
