LOCUS       sequence-444947-        6019 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 6019)
  AUTHORS   Tong Q, Tsai J, Tan G, Dalgin G, Hotamisligil GS
  TITLE     Interaction between GATA and the C/EBP family of transcription 
            factors is critical in GATA-mediated suppression of adipocyte 
            differentiation.
  JOURNAL   Mol Cell Biol 2005 Jan;25(2):706-15.
  PUBMED    15632071
REFERENCE   2  (bases 1 to 6019)
  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: "Mol Cell 
            Biol"; date: "2005-01"; volume: "25(2)"; pages: "706-15"
COMMENT     Sequence Label: pFLAG-GATA3
FEATURES             Location/Qualifiers
     source          1..6019
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        128..331
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     282..302
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             365..388
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1790..2412
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1989..2008)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2436..2456)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2441..2770
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2621..2756
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2808..2827)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2809..2827)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2841..2857)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2841..2857)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2854..2876)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2865..2881
                     /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(2889..2895,2896..2913,2914..2919))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2934..2955
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3072..3089)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3243..3831)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3323..3342)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4002..4793,4794..4862))
                     /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     4625..4644
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4863..4967)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4994..5449
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5081..5100)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5291..5312
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5575..5597
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5589..5606
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5590..5606
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        join(5767..6019,1..127)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
       61 gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      121 taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt ttgactcacg
      181 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      241 acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg
      301 tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga attgatctac
      361 catggactac aaagacgatg acgacaagct tgcggccgcg aattcatcga tagatctgat
      421 cgaggtgact gcggaccagc cgcgctgggt gagccaccat caccccgcgg tcctcaacgg
      481 tcagcaccca gacacgcacc acccgggcct cggccattcg tacatggaag ctcagtatcc
      541 gctgacggaa gaggtggacg tactttttaa catcgatggt caaggcaacc acgtcccgtc
      601 ctactacgga aactccgtca gggctacggt gcagaggtat cctccgaccc accacgggag
      661 ccaggtatgc cgcccgcctc tgctgcacgg atctctgccc tggctggatg gcggcaaagc
      721 cctgagcagc caccacaccg cctcgccctg gaacctcagc cccttctcca agacgtccat
      781 ccaccacggc tctccggggc ctctgtccgt ttaccctccg gcttcatcct cttctctggc
      841 ggccggccac tccagtcctc atctcttcac cttcccgccc accccgccga aagacgtctc
      901 cccagacccg tcgctgtcca ccccgggatc cgccgggtcg gccaggcaag atgagaaaga
      961 gtgcctcaag tatcaggtgc agctgccaga tagcatgaag ctggagacgt ctcactctcg
     1021 aggcagcatg accaccctgg gtggggcctc atcctcagcc caccacccca ttaccaccta
     1081 tccgccctat gtgcccgagt acagctctgg actcttccca cccagcagcc tgctgggagg
     1141 atcccctacc gggttcggat gtaagtcgag gcccaaggca cgatccagca cagaaggcag
     1201 ggagtgtgtg aactgcgggg caacctctac cccactgtgg cggcgagatg gtaccgggca
     1261 ctacctttgc aatgcctgcg gactctacca taaaatgaat gggcagaacc ggccccttat
     1321 caagcccaag cgaaggctgt cggcagcaag gagagcaggg acatcctgcg cgaactgtca
     1381 gaccaccacc accaccctct ggaggaggaa cgctaatggg gacccggtct gcaatgcctg
     1441 tgggctgtac tacaagcttc ataatattaa cagacccctg actatgaaga aagaaggcat
     1501 ccagacccga aaccggaaga tgtctagcaa atcgaaaaag tgcaaaaagg tgcatgacgc
     1561 gctggaggac ttccccaaga gcagctcctt caacccggcc gctctctcca gacacatgtc
     1621 atccctgagc cacatctctc ccttcagcca ctccagccac atgctgacca caccgacgcc
     1681 catgcatccg ccctccggcc tctccttcgg acctcaccac ccttccagca tggtcaccgc
     1741 catgggttag agaggcatcg atcggtacca gtcgactcta gaggatcccg ggtggcatcc
     1801 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     1861 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     1921 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     1981 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     2041 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     2101 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     2161 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     2221 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttaaaata
     2281 actataccag caggaggacg tccagacaca gcataggcta cctggccatg cccaaccggt
     2341 gggacatttg agttgcttgc ttggcactgt cctctcatgc gttgggtcca ctcagtagat
     2401 gcctgttgaa ttgggtacgc ggccagcttg gctgtggaat gtgtgtcagt tagggtgtgg
     2461 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2521 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2581 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     2641 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     2701 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     2761 cttttgcaaa aagctcctcg aggaactgaa aaaccagaaa gttaattccc tatagtgagt
     2821 cgtattaaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     2881 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     2941 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3001 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3061 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3121 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3181 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3241 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3301 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3361 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3421 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3481 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3541 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3601 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3661 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3721 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3781 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3841 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3901 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3961 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4021 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4081 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4141 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4201 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4261 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4321 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4381 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4441 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4501 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4561 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4621 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4681 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4741 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4801 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4861 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4921 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4981 aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5041 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     5101 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     5161 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     5221 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     5281 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     5341 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     5401 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat
     5461 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     5521 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     5581 cacgacgttg taaaacgacg gccagtgcca agctgatcta tacattgaat caatattggc
     5641 aattagccat attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc
     5701 atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc
     5761 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     5821 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     5881 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     5941 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     6001 tacatcaagt gtatcatat
//
