LOCUS       sequence-301960-        6185 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 6185)
  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 6185)
  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-GATA2
FEATURES             Location/Qualifiers
     source          1..6185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..45
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     37..54
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     38..54
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        215..594
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        595..798
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     749..769
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             832..855
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2423..3045
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2622..2641)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3069..3089)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3074..3403
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3254..3389
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3441..3460)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3442..3460)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3474..3490)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3474..3490)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3487..3509)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3498..3514
                     /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(3522..3528,3529..3546,3547..3552))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3567..3588
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3705..3722)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3876..4464)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3956..3975)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4635..5426,5427..5495))
                     /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     5258..5277
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5496..5600)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5627..6082
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5714..5733)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5924..5945
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag
       61 ctgatctata cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca
      121 taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt
      181 tatattggct catgtccaat atgaccgcca tgttgacatt gattattgac tagttattaa
      241 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      301 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      361 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      421 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc
      481 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      541 cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      601 cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      661 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      721 aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt acggtgggag
      781 gtctatataa gcagagctcg tttagtgaac cgtcagaatt gatctaccat ggactacaaa
      841 gacgatgacg acaagcttgc ggccgcgaat tcatcgatag atctgatcga ggtggcgccc
      901 gagcagccgc gctggatggc gcacccggcc gtgctgaatg cgcagcaccc cgactcacac
      961 cacccgggcc tggcgcacaa ctacatggaa cccgcgcagc tgctgcctcc agacgaggtg
     1021 gacgtcttct tcaatcacct cgactcgcag ggcaacccct actatgccaa ccccgctcac
     1081 gcgcgggcgc gcgtctccta cagccccgcg cacgcccgcc tgaccggagg ccagatgtgc
     1141 cgcccacact tgttgcacag cccgggtttg ccctggctgg acgggggcaa agcagccctc
     1201 tctgccgctg cggcccacca ccacaacccc tggaccgtga gccccttctc caagacgcca
     1261 ctgcacccct cagctgctgg aggccctgga ggcccactct ctgtgtaccc aggggctggg
     1321 ggtgggagcg ggggaggcag cgggagctca gtggcctccc tcacccctac agcagcccac
     1381 tctggctccc accttttcgg cttcccaccc acgccaccca aagaagtgtc tcctgaccct
     1441 agcaccacgg gggctgcgtc tccagcctca tcttccgcgg ggggtagtgc agcccgagga
     1501 gaggacaagg acggcgtcaa gtaccaggtg tcactgacgg agagcatgaa gatggaaagt
     1561 ggcagtcccc tgcgcccagg cctagctact atgggcaccc agcctgctac acaccacccc
     1621 atccccacct acccctccta tgtgccggcg gctgcccacg actacagcag cggactcttc
     1681 caccccggag gcttcctggg gggaccggcc tccagcttca cccctaagca gcgcagcaag
     1741 gctcgttcct gttcagaagg ccgggagtgt gtcaactgtg gggccacagc cacccctctc
     1801 tggcggcggg acggcaccgg ccactacctg tgcaatgcct gtggcctcta ccacaagatg
     1861 aatgggcaga accgaccact catcaagccc aagcgaagac tgtcggccgc cagaagagcc
     1921 ggcacctgtt gtgcaaattg tcagacgaca accaccacct tatggcgccg aaacgccaac
     1981 ggggaccctg tctgcaacgc ctgtggcctc tactacaagc tgcacaatgt taacaggcca
     2041 ctgaccatga agaaggaagg gatccagact cggaaccgga agatgtccaa caagtccaag
     2101 aagagcaaga aaggggcgga gtgcttcgag gagctgtcaa agtgcatgca ggagaagtca
     2161 tcccccttca gtgcagctgc cctggctgga cacatggcac ctgtgggcca cctcccgccc
     2221 ttcagccact ccggacacat cctgcccact ccgacgccca tccacccctc ctccagcctc
     2281 tccttcggcc acccccaccc gtccagcatg gtgaccgcca tgggctaggg aacagatgga
     2341 cgtcgaggac cgggcactcc cgggatgggt ggaccaaacc cttaccagca tcgatcggta
     2401 ccagtcgact ctagaggatc ccgggtggca tccctgtgac ccctccccag tgcctctcct
     2461 ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat
     2521 cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag
     2581 caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg
     2641 gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct
     2701 cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt
     2761 tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta
     2821 atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac
     2881 tgctcccttc cctgtccttc tgattttaaa ataactatac cagcaggagg acgtccagac
     2941 acagcatagg ctacctggcc atgcccaacc ggtgggacat ttgagttgct tgcttggcac
     3001 tgtcctctca tgcgttgggt ccactcagta gatgcctgtt gaattgggta cgcggccagc
     3061 ttggctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3121 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3181 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3241 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3301 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3361 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc tcgaggaact
     3421 gaaaaaccag aaagttaatt ccctatagtg agtcgtatta aattcgtaat catgtcatag
     3481 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3541 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3601 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3661 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3721 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3781 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3841 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3901 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3961 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4021 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4081 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4141 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4201 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4261 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4321 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4381 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4441 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4501 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4561 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4621 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4681 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4741 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4801 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4861 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4921 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4981 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5041 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5101 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5161 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5221 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5281 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5341 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5401 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5461 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5521 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5581 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc
     5641 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     5701 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     5761 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     5821 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     5881 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     5941 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     6001 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     6061 aaaatattaa cgcttacaat ttccattcgc cattcaggct gcgcaactgt tgggaagggc
     6121 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     6181 gatta
//
