LOCUS       sequence-227505-        6721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid expresses human GATA-1 K137R mutant. This construct 
            cannot undergo sumoylation on K137..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6721)
  AUTHORS   Hietakangas V, Anckar J, Blomster HA, Fujimoto M, Palvimo JJ, Nakai 
            A, Sistonen L
  TITLE     PDSM, a motif for phosphorylation-dependent SUMO modification.
  JOURNAL   Proc Natl Acad Sci U S A. 2006 Jan 3;103(1):45-50. doi: 
            10.1073/pnas.0503698102. Epub 2005 Dec 21.
  PUBMED    16371476
REFERENCE   2  (bases 1 to 6721)
  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"; doi: 
            "10.1073/pnas.0503698102"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2006-01-3- 3"; volume: "103"; issue: "1"; pages: "45-50"
COMMENT     Sequence Label: hGATA-1 K137R
FEATURES             Location/Qualifiers
     source          1..6721
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1972..2001
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2017..2034
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2078..2095)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2084..2308
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2354..2782
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2441..2460)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2651..2672
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2791..2811)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2796..3125
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2976..3111
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3038..3057
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3192..3986
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3246..3265)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3856..3875
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4160..4281
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4197..4216)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4251..4270
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4330..4346)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4330..4346)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4343..4365)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4354..4370
                     /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(4378..4384,4385..4402,4403..4408))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4423..4444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4561..4578)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4732..5320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4812..4831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5491..6282,6283..6351))
                     /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     6114..6133
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6352..6456)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6531..6550)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacgggccc tctagactcg agcggccgca tggagttccc tggcctgggg
      721 tccctgggga cctcagagcc cctcccccag tttgtggatc ctgctctggt gtcctccaca
      781 ccagaatcag gggttttctt cccctctggg cctgagggct tggatgcagc agcttcctcc
      841 actgccccga gcacagccac cgctgcagct gcggcactgg cctactacag ggacgctgag
      901 gcctacagac actccccagt ctttcaggtg tacccattgc tcaactgtat ggaggggatc
      961 ccagggggct caccatatgc cggctgggcc tacggcaaga cggggctcta ccctgcctca
     1021 actgtgtgtc ccacccgcga ggactctcct ccccaggccg tggaagatct ggatggaaaa
     1081 ggcagcacca gcttcctgga gactttgagg acagagcggc tgagcccaga cctcctgacc
     1141 ctgggacctg cactgccttc atcactccct gtccccaata gtgcttatgg gggccctgac
     1201 ttttccagta ccttcttttc tcccaccggg agccccctca attcagcagc ctattcctct
     1261 cccaagcttc gtggaactct ccccctgcct ccctgtgagg ccagggagtg tgtgaactgc
     1321 ggagcaacag ccactccact gtggcggagg gacaggacag gccactacct atgcaacgcc
     1381 tgcggcctct atcacaagat gaatgggcag aacaggcccc tcatccggcc caagaagcgc
     1441 ctgattgtca gtaaacgggc aggtactcag tgcaccaact gccagacgac caccacgaca
     1501 ctgtggcgga gaaatgccag tggggatccc gtgtgcaatg cctgcggcct ctactacaag
     1561 ctacaccagc ctcctttttg gcaggtgaac cggccactga ccatgcggaa ggatggtatt
     1621 cagactcgaa accgcaaggc atctggaaaa gggaaaaaga aacggggctc cagtctggga
     1681 ggcacaggag cagccgaagg accagctggt ggctttatgg tggtggctgg gggcagcggt
     1741 agcgggaatt gtggggaggt ggcttcaggc ctgacactgg gccccccagg tactgcccat
     1801 ctctaccaag gcctgggccc tgtggtgctg tcagggcctg ttagccacct catgcctttc
     1861 cctggacccc tactgggctc acccacgggc tccttcccca caggccccat gccccccacc
     1921 accagcacta ctgtggtggc tccgctcagc tcaggtacca agcttgggcc cgaacaaaaa
     1981 ctcatctcag aagaggatct gaatagcgcc gtcgaccatc atcatcatca tcattgagtt
     2041 taaacggtct ccagcttaag tttaaaccgc tgatcagcct cgactgtgcc ttctagttgc
     2101 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2161 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2221 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2281 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     2341 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2401 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2461 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2521 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     2581 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2641 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     2701 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     2761 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     2821 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2881 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2941 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3001 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3061 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3121 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     3181 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3241 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3301 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3361 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3421 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3481 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3541 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3601 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3661 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     3721 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3781 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3841 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3901 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     3961 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4021 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4081 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4141 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4201 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4261 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     4321 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4381 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4441 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4501 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4561 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4621 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4681 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4741 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4801 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4861 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4921 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4981 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5041 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5101 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5161 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5221 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5281 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5341 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5401 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5461 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5521 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5581 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5641 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5701 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5761 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5821 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5881 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5941 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6001 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6061 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6121 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6181 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6241 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6301 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6361 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6421 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6481 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     6541 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     6601 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     6661 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
     6721 t
//
