LOCUS       sequence-6862-ex        7962 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7962)
  AUTHORS   Malik TH, Shoichet SA, Latham P, Kroll TG, Peters LL, Shivdasani RA
  TITLE     Transcriptional repression and developmental functions of the 
            atypical vertebrate GATA protein TRPS1.
  JOURNAL   EMBO J. 2001 Apr 2. 20(7):1715-25.
  PUBMED    11285235
REFERENCE   2  (bases 1 to 7962)
  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: "EMBO J. 
            2001 Apr 2. 20(7):1715-25."
COMMENT     Sequence Label: p061 pCS xTRPS1
FEATURES             Location/Qualifiers
     source          1..7962
                     /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    4009..4143
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4033..4052)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4087..4106
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4260..4280)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4260..4278)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4299..4315)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4299..4315)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4312..4334)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4323..4339
                     /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(4347..4353,4354..4371,4372..4377))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4392..4413
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4530..4547)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4701..5289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4781..4800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5460..6251,6252..6320))
                     /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     6083..6102
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6321..6425)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6451..6906)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6588..6609)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6800..6819
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6978..7962
                     /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 ttcgaattcg tcgggggtgg tgccttcctc
      121 tgcttaaggc tcctttgata ttaatagtgt tggtgtcttg aaactgacgt aatgctggag
      181 actgaggtcc tgacaagcga taacatttct gataaagaac tcatctcgct gcaatctttg
      241 tgtgattcct ccagagccac agaatctcct gtttagagcc acagcatcca gattttcttt
      301 tctctctctc tctctccctc caccccttta acaacacccc tctccttgtt cctgtctggg
      361 tctctgctct cagccctaca cacttggggg ggaacaggaa gccttttctt ctcccccccc
      421 ttcaaacaca aaaataatca aagatcaact cggcaaacaa caaggaaacg gttcatggct
      481 cacgccacag ttccaccatc ttccagactg ctgctgctgc tgctaaggtt attgcagatc
      541 aatcaacagc actgacacta tactctgccc tttacaaaat gagatacaac gttggttaaa
      601 caaaggcaaa gcttcgacac aaggatatgg tccggaaaaa gaacccccct ctgagaaaca
      661 ttgcaagtga aggtgaggca cagataacag agtctgctgc atccaagaga gaagacacta
      721 ttagcagcaa ggaaatttcc actgatccga tgcaagaaaa tagtgaacaa agtggcttgg
      781 tggaacacaa cagtgatgac cacagctttc atgatcaaga gccttcctcc agcatcaaca
      841 aggactctga aagcttgtca ttaagtgaaa gagctgtagt caactacagc caccttaagg
      901 gaagaaatgt ttatttttct ccgatggagg tgaccgacag aaacatgtta gcattagtca
      961 ccacagatac tagatctgct tgtgatcctc ttaagtcccc aataaaatca gaagctgatg
     1021 acacccaaga actagcctca tctgcatctg tggacagttt agaggccaaa gaggaaaatg
     1081 atatgtcacc aagggctact gacttcacag tgcaatgtgg caaagtagac tgtcaaagct
     1141 caagtccagc ttcggtggcc tcagataacc tacatgtgcc ttcagatggg attgcggggc
     1201 tgaacaaatc tcaagctgtt ctgctggtca atgacaattc tgattcagca cctttatccc
     1261 cagagcttca ggattttaaa tgcaacattt gtgggtatgg atactatgga aatgacccca
     1321 cagaccttat taaacacttc cgtaaatacc atctcggcct gcacaaccga accaggcaag
     1381 atgttgaact tgacaccaaa atcctggcac ttcacaatat ggtacaattt agtcaatcca
     1441 aagatttcca gaagatgaat cgttctgtgc tttcaggggt tttgcaggac ttcaactctc
     1501 cgagacctgt tctgctaaat ggaacatatg acgtgcaggt aacctttggt gaaacattta
     1561 ttggcattgg gcgtaaaact ccagactgcc aaggaaacac caagtacttt cgctgcaaat
     1621 tctgcaactt cacgtacatg ggcaactcgg ccacagagtt ggagcaacac tttcttaaga
     1681 cgcatcctaa caaaatgaaa atgtcctctg attctgatgt cctcctgctt cattatgaga
     1741 atgcacatga agcccaggca tgtgagatca agcaggaact gaatcaccag catggggccg
     1801 atgggcagcc gagcatcaaa gaaatcaaag aacactcatg taccaaatgt gactttattg
     1861 tccaggtgga agaagatctg ccccgacact acaggagggt gcataactgc tacaaatgtc
     1921 gtcagtgcaa ctttacagca gcagacaccc agtcactatt ggatcacttt aactctgctc
     1981 attgtcagga gtttgaaatc actacatcta atggagggga gcatcatgga acatcttcta
     2041 ttaaggaaga accaaaaact gatctaaaag tgtataatct tgtcactcct gattccaaaa
     2101 tgggagaagc catatttgac agcactgtga aaaaagagaa gcttgaggat aaggaaacac
     2161 tcagggagaa agcttggtct gatggttctg ttgatgatct tcggggtgtt gcctggagag
     2221 caccagacat tttaagaaca agcccatctt actcacagat gggcttgggt cttttaacca
     2281 cagtgtctgt aaaccaagat cagcaaaaat cctcaagaga cagccccaat gtggaagcag
     2341 cacatctagc caggccagtg tatggcttgt ctatagaacc caagggattt caaggagtga
     2401 cagcaggttc tagtggagaa aggtctggtc agcatacacc acagtatcct acagcaggag
     2461 atagcaaatc aaaagatgaa tcccagtcac tgctaaggag gcgtagagga tccggtgtgt
     2521 tctgtgccaa ctgtctgacc acaaagacat ccctctggag gaaaaatgcc aacggtggct
     2581 atgtctgcaa tgcgtgtggc ctttaccaga aacttcactc gactcccaga ccgttaaata
     2641 taattaagca gaacaatggt gagcagatca ttaggagacg aacaagaaag cgccttaacc
     2701 cagaagcact gcagcctgaa cagttaacta aacatcaaag agcgagtagt gaagaacaag
     2761 ctaatggaag tcctttagat ataaggtcag aggaccactc aatggaaggt catcaaagag
     2821 aaaatcagca gcttagtatg aataaatatg ggtctcaagc ttcattgact aaaagccatt
     2881 ctgctcagca gaccatgatt gtcagtcaaa ccatggatat acacaaacgg atgcaacctt
     2941 tgcacattca gataaaaagc cctcaggaaa gcagtggaga accaggaaac agctcatctg
     3001 tttctgatgg caaaggaagc tcagaaagag gaagtccaat tgagaaatac atgagaccta
     3061 taaagcatcc taattactct ccacctggaa gcccaataga aaaatatcag tacccactct
     3121 ttggacttcc ttttgttcac aatgacttcc agagtgaagc tgactggctg agattctgga
     3181 gtaaatataa actgtcagtt cctggaaatc cgcactactt gagtcatgtg cctggcctgc
     3241 caaatccctg cccaaattat gtgccttatc ccacctttaa tctgcctgct caatattcct
     3301 ctgttgggtc agacaatgac attcctctag atttggcaat gaagcattcc agacctggat
     3361 caggaacaaa tggagactct aaggaaaaat caaagtcacc cgtgagtgta aaggatgatg
     3421 gtcctttgaa tgtaactaaa atagagaaat cagataagag tacgcaagat gaactttcaa
     3481 caaaatgtgt gcactgtggc attgtctttc tggatgaggt tatgtatgct ctacatatga
     3541 gttgccatgg ggaaagtgga ccttttcagt gtagtatatg ccaacatctt tgcacggaca
     3601 agtatgactt cacaactcat attcagagag gcctacatag gaacattgca caagctgaaa
     3661 agaatggaaa aaataaagat taaaacttta gcacttagca caatttagta gaaatggggt
     3721 ttcttgaata gaaattcagt agcttgtgac ttctaatgag ggccagtaaa gcacttccta
     3781 aagcatgttc aaagactcat gttctccgta actaaaactt gtcaaatatt agccatacgt
     3841 aagcagcatt tccagtataa aaaaagtgca gatatggggt gccttgatct atggttttca
     3901 ggaaaaggaa tgcctgaaaa cttaataaag tattaaaaaa ggaaaaaaaa aaaaaagcgg
     3961 ccgcaaggcc tctcgagcct ctagaactat agtgagtcgt attacgtaga tccagacatg
     4021 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     4081 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     4141 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     4201 ttttaattcg cggcgcgccg cggcgccaat gcattgggcc cggtacccag cttttgttcc
     4261 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     4321 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4381 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4441 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4501 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4561 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4621 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4681 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4741 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4801 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4861 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4921 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4981 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5041 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5101 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     5161 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5221 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5281 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5341 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5401 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5461 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5521 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5581 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5641 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5701 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5761 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5821 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5881 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5941 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6001 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6061 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6121 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6181 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6241 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6301 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6361 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6421 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     6481 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     6541 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     6601 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     6661 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     6721 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     6781 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     6841 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     6901 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     6961 cttcgctatt acgccaggac catagccaat tcaatatggc gtatatggac tcatgccaat
     7021 tcaatatggt ggatctggac ctgtgccaat tcaatatggc gtatatggac tcgtgccaat
     7081 tcaatatggt ggatctggac cccagccaat tcaatatggc ggacttggca ccatgccaat
     7141 tcaatatggc ggacctggca ctgtgccaac tggggagggg tctacttggc acggtgccaa
     7201 gtttgaggag gggtcttggc cctgtgccaa gtccgccata ttgaattggc atggtgccaa
     7261 taatggcggc catattggct atatgccagg atcaatatat aggcaatatc caatatggcc
     7321 ctatgccaat atggctattg gccaggttca atactatgta ttggccctat gccatatagt
     7381 attccatata tgggttttcc tattgacgta gatagcccct cccaatgggc ggtcccatat
     7441 accatatatg gggcttccta ataccgccca tagccactcc cccattgacg tcaatggtct
     7501 ctatatatgg tctttcctat tgacgtcata tgggcggtcc tattgacgta tatggcgcct
     7561 cccccattga cgtcaattac ggtaaatggc ccgcctggct caatgcccat tgacgtcaat
     7621 aggaccaccc accattgacg tcaatgggat ggctcattgc ccattcatat ccgttctcac
     7681 gccccctatt gacgtcaatg acggtaaatg gcccacttgg cagtacatca atatctatta
     7741 atagtaactt ggcaagtaca ttactattgg aagtacgcca gggtacattg gcagtactcc
     7801 cattgacgtc aatggcggta aatggcccgc gatggctgcc aagtacatcc ccattgacgt
     7861 caatggggag gggcaatgac gcaaatgggc gttccattga cgtaaatggg cggtaggcgt
     7921 gcctaatggg aggtctatat aagcaatgct cgtttaggga ac
//
