LOCUS       sequence-248960-        6517 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 6517)
  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 6517)
  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-GR
FEATURES             Location/Qualifiers
     source          1..6517
                     /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    2569..2703
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2593..2612)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2647..2666
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2816..2836)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2816..2834)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2855..2871)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2855..2871)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2868..2890)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2879..2895
                     /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(2903..2909,2910..2927,2928..2933))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2948..2969
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3086..3103)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3257..3845)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3337..3356)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4016..4807,4808..4876))
                     /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     4639..4658
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4877..4981)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5007..5462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5144..5165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5356..5375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5533..6517
                     /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 ttcgaattca aggtcccttg ccagtgccac
      121 tgcctgaagt gcaggctccc actggaatat ggcggacaca gaggaggctt atgggatgcc
      181 tgatacccct gtggaagcag agcccaaaga actacagtgt gagcctaaac aggacaatca
      241 attgggggca agcagcaaga cccccacttc tcctccggct gcatttaccc agcagggaat
      301 ggaaggcatt aaggtgtttt tgcatgagag agaactatgg ctgaaatttc atgaagttgg
      361 cacagaaatg attattacta aggctggaag gcgcatgttc cctagttaca aagtgaaggt
      421 tactggcctg aacccaaaaa ccaagtacat cctcctcatg gatatcgttc ctgcagatga
      481 ccacagatac aagtttgctg ataacaaatg gtcagtcaca ggcaaggctg aacccgctat
      541 gccaggacgt ttgtatgttc acccagattc accagccacg ggcgcacatt ggatgaggca
      601 gctagtctcc ttccagaagc tgaagctcac aaacaaccat ctggatccat ttggacatat
      661 catcctgaat tcaatgcaca agtaccagcc acggctccac attgtcaagg cagatgagaa
      721 caatggcttt gggtccaaga atacagcttt ctgcactcat gtattttctg agactgattt
      781 tattgctgtc acctcatatc agaaccacaa gatcacacag ctaaagatag agaacaatcc
      841 ctttgccaaa ggatttcggg gaagtgatga catggagcta caccggatgt ccaggatgca
      901 gagcaaagag tatccagtag tgccaagaag cacagtccgt cagaaagtct cctctaatca
      961 cagcccattc tctcaagaaa cacgtaacat tacaggatct tctaccctaa attcccaata
     1021 tcagtgtgag aatggtgtta gcagcacatc tcaggatctt ctgccatctt ccagtgcata
     1081 tacttccctg ccccatgagt ctggtactat atatcactgc acaaaaagaa aagtaagtga
     1141 agagccagct gagcattcct ataagaaacc ctacatggac acttcaccaa gcgaagagga
     1201 ccctttttac cgatctgggt acccacaacc ttcatcctct tcctcatcta ccacctcttt
     1261 ccgcacagaa tctgcccaac gacaggcctg catgtatgct agttcagcac ctgccacaga
     1321 gcctgtacca tccatagagg atatatcttg caacagctgg tctagtgtgc cctcttacag
     1381 cagctgtaca gtaggtggag gaatgcaacc catggagcgc ctcccatatc agcacttctc
     1441 tgctcacttt acctctagtt ccctcatgcc tagactgagt aatcatgcag gcacacagcc
     1501 aagtgattct cactccatgt tccaacatca gtcttctcat caggccattg taaggcaatg
     1561 caatcctcaa tctgggttac aacaatcatc agctcttcag cccactgaat tcttgtaccc
     1621 acacagtgtt ccccggacca tttcaccaca tcaataccac tctgtgcatg gggtaggtat
     1681 ggtcccagac tcgcctctcg agcctctagg gcgccgctcg agcccctctg aaaatcctgg
     1741 taacaaaaca atagttcctg caacgttacc acaactcacc cctaccctgg tgtcactgtt
     1801 ggaggttatt gaacctgaag tgttatatgc aggatatgat agctctgttc cagactcaac
     1861 ttggaggatc atgactacgc tcaacatgtt aggagggcgg caagtgattg cagcagtgaa
     1921 atgggcaaag gcaataccag gtttcaggaa cttacacctg gatgaccaaa tgaccctact
     1981 gcagtactcc tggatgtttc ttatggcatt tgctctgggg tggagatcat atagacaatc
     2041 aagtgcaaac ctgctgtgtt ttgctcctga tctgattatt aatgagcaga gaatgactct
     2101 accctgcatg tacgaccaat gtaaacacat gctgtatgtt tcctctgagt tacacaggct
     2161 tcaggtatct tatgaagagt atctctgtat gaaaacctta ctgcttctct cttcagttcc
     2221 taaggacggt ctgaagagcc aagagctatt tgatgaaatt agaatgacct acatcaaaga
     2281 gctaggaaaa gccattgtca agagggaagg aaactccagc cagaactggc agcggtttta
     2341 tcaactgaca aaactcttgg attctatgca tgaagtggtt gaaaatctcc ttaactattg
     2401 cttccaaaca tttttggata agaccatgag tattgaattc cccgagatgt tagctgaaat
     2461 catcaccaat cagataccaa aatattcaaa tggaaatatc aaaaaacttc tgtttcatca
     2521 aaagtgctcg agtgcccgcc ctagaactat agtgagtcgt attacgtaga tccagacatg
     2581 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     2641 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     2701 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     2761 ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt
     2821 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     2881 gttatccgct cacaattcca cacaacatac gagccgggag cataaagtgt aaagcctggg
     2941 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     3001 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     3061 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3121 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3181 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3241 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3301 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3361 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3421 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3481 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3541 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3601 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3661 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3721 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3781 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3841 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3901 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3961 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4021 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4081 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4141 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4201 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4261 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4321 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4381 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4441 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4501 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4561 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4621 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4681 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4741 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4801 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4861 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4921 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4981 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     5041 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     5101 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     5161 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     5221 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     5281 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     5341 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     5401 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     5461 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     5521 gctattacgc cagtcgaccg ccaattcaat atggcgtata tggactcatg ccaattcaat
     5581 atggtggatc tggacctgtg ccaattcaat atggcgtata tggactcgtg ccaattcaat
     5641 atggtggatc tggaccccag ccaattcaat atggcggact tggcaccatg ccaattcaat
     5701 atggcggacc tggcactgtg ccaactgggg aggggtctac ttggcacggt gccaagtttg
     5761 aggaggggtc ttggccctgt gccaagtccg ccatattgaa ttggcatggt gccaataatg
     5821 gcggccatat tggctatatg ccaggatcaa tatataggca atatccaata tggccctatg
     5881 ccaatatggc tattggccag gttcaatact atgtattggc cctatgccat atagtattcc
     5941 atatatgggt tttcctattg acgtagatag cccctcccaa tgggcggtcc catataccat
     6001 atatggggct tcctaatacc gcccatagcc actcccccat tgacgtcaat ggtctctata
     6061 tatggtcttt cctattgacg tcatatgggc ggtcctattg acgtatatgg cgcctccccc
     6121 attgacgtca attacggtaa atggcccgcc tggctcaatg cccattgacg tcaataggac
     6181 cacccaccat tgacgtcaat gggatggctc attgcccatt catatccgtt ctcacgcccc
     6241 ctattgacgt caatgacggt aaatggccca cttggcagta catcaatatc tattaatagt
     6301 aacttggcaa gtacattact attggaagta cgccagggta cattggcagt actcccattg
     6361 acgtcaatgg cggtaaatgg cccgcgatgg ctgccaagta catccccatt gacgtcaatg
     6421 gggaggggca atgacgcaaa tgggcgttcc attgacgtaa atgggcggta ggcgtgccta
     6481 atgggaggtc tatataagca atgctcgttt agggaac
//
