LOCUS       sequence-463062-        6130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV production plasmid for EPOR(synEPOR) vector from Figs. 3-5 that 
            mediates HDR at EPOR locus using EPOR gRNA. synEPOR is codon 
            diverged to prevent premature recombination with EPOR locus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6130)
  AUTHORS   Shah AP, Majeti KR, Ekman FK, Selvaraj S, Soupene E, Chati P, Sinha 
            R, Luna SE, Charlesworth CT, McCreary T, Lesch BJ, Tran T, Sharma D,
            Chu SN, Porteus MH, Cromer MK
  TITLE     Engineering inducible signaling receptors to enable 
            erythropoietin-free erythropoiesis
  JOURNAL   bioRxiv 2024.04.12.589260
REFERENCE   2  (bases 1 to 6130)
  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: "bioRxiv 
            2024.04.12.589260"
COMMENT     Sequence Label: MKC176_EPOR(2A-synEPOR)
FEATURES             Location/Qualifiers
     source          1..6130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   927..1056
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(1118..1706)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1198..1217)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1877..2668,2669..2737))
                     /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     2500..2519
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2738..2842)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2910..2928
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2966..2988)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3088..3107
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3124..3579)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3261..3282)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3473..3492
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3654..3794
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3665..3794
                     /label=AAV2 ITR
     CDS             4791..4844
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4956..5276
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    5862..6086
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 cctgtacctt gtggtatctg actctggcat ctcaactgac tacagctcag gggactccca
       61 gggagcccaa gggggcttat ccgatggccc ctactccaac ccttatgaga acagccttat
      121 cccagccgct gagcctctgc cccccagcta tgtggcttgc tcttaggaca ccaggctgca
      181 gatgatcagg gatccaatat gactcagaga accagtgcag actcaagact tatggaacag
      241 ggatggcgag gcctctctca ggagcagggg cattgctgat tttgtctgcc caatccatcc
      301 tgctcaggaa accacaacct tgcagtattt ttaaatatgt atagtttttt tttgtatcta
      361 tatatatata tacacatatg tatgtaagtt tttctaccat gatttctaca aacacccttt
      421 aagtcccatc ttcccctggg cataggccat agggatagaa gttaaagttc ttgagcttat
      481 tcagaagctg gatctgcaat ctgaatgcta ctcataacat aacaaaatag tatgttaaac
      541 agctcttaaa tcttactggc ttaccacatt aaatgatttc tctctcctaa ctcagctcaa
      601 atgggcagcc atccatggga tgagtcagag gttcagactc ttccagtctg tagctctacc
      661 ttctcttagg gtacttagat ggatcccctg ttctacaaac tgccagtcag caagggaaga
      721 aaaagggcag caatgaccct caatgggcca tttgagggat ctggcctgga aatgggcttc
      781 ctctcttctt ctcacacctc actggctgga aacagtcaca tgaccccagt cacatgaaag
      841 gccaggaaac ttagtttagc tgtacaccca ggaagggcaa agctgtttaa gggccactag
      901 ctagtctctg ccactaatgc ggccgcagga acccctagtg atggagttgg ccactccctc
      961 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg
     1021 cctcagtgag cgagcgagcg cgcagctgcc tgcaggacat gtgagcaaaa ggccagcaaa
     1081 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     1141 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     1201 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1261 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     1321 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1381 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1441 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1501 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     1561 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     1621 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     1681 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     1741 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     1801 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     1861 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     1921 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     1981 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     2041 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     2101 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     2161 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     2221 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     2281 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     2341 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     2401 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     2461 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     2521 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     2581 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     2641 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     2701 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     2761 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     2821 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     2881 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     2941 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     3001 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     3061 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     3121 ataaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3181 cattttttaa ccaatagacc gaaatcggca aaatccctta taaatcaaaa gaatagcccg
     3241 agatagagtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3301 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3361 ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3421 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3481 aagcgaaagg agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3541 ccacacccgc cgcgcttaat gcgccgctac agggcgcgta ctatggttgc tttgacgtat
     3601 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gcccctgcag
     3661 gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac
     3721 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gaaggagtgg ccaactccat
     3781 cactaggggt tcctgcggcc gcgagatggg ggtctcacta tgtttcctag gctggtcttg
     3841 aactcctggg ttcaaatgat cctcccacct cagcctccca aagtactggg attatagtgc
     3901 tggtgtaaac cactgcacct ggccatggcc aggattaaag ggagaatgac caaggtatat
     3961 tgaactccta tgcacccttc aataccctgt tccatttacc cttttgtagg gccttgctga
     4021 tgcttcagcc aaaacccctg tcccctggcc ctgatgtact cctctgcctc cattgtgatc
     4081 acagggacca agtgtatctg tgcctctatg actgggagtg gagggggaat tggtgagtat
     4141 tcaatgagtc atatctatgt aactatttat attggcttca acagggctct gaagcagaag
     4201 atctggcctg gcatcccgag cccagagagc gagtttgaag gcctcttcac cacccacaag
     4261 ggtaacttcc aggtaggtgg cctggttgtc ccctcagtgc ctgggcttcc ctgcttcttg
     4321 cagccaaact gcaggcctct ctgagcaggt tggtgctatt tcttcagctg tggctgtacc
     4381 agaatgatgg ctgcctgtgg tggagcccct gcaccccctt cacggaggac ccacctgctt
     4441 ccctggaagt cctctcagag cgctgctggg ggacgatgca ggcagtggag ccggggacag
     4501 atgatgaggg ccccctgctg gagccagtgg gcagtgagca tgcccaggat acctatctgg
     4561 tgctggacaa atggttgctg ccccggaacc cgcccagtga ggacctccca gggcctggtg
     4621 gcagtgtgga catagtggcc atggatgaag gctcagaagc atcctcctgc tcatctgctt
     4681 tggcctcgaa gcccagccca gagggagcct ctgctgccag ctttgagtac actatcctgg
     4741 accccagctc ccagctcttg cgtccacgca aaagacgctc cggttctgga gagggcaggg
     4801 ggagtcttct tacatgcggg gatgttgagg agaatcccgg acccatgaac tccttctcca
     4861 caagcgcctt cggtccagtt gccttctccc tggggctgct cctggtgttg cctgctgcct
     4921 tccctgcccc aggcggcggc ggaagcatgc ttgagggagt gcaggtggaa accatctccc
     4981 caggagacgg gcgcaccttc cccaagcgcg gccagacctg cgtggtgcac tacaccggga
     5041 tgcttgaaga tggaaagaaa gttgattcct cccgggacag aaacaagccc tttaagttta
     5101 tgctaggcaa gcaggaggtg atccgaggct gggaagaagg ggttgcccag atgagtgtgg
     5161 gtcagagagc caaactgact atatctccag attatgccta tggtgccact gggcacccag
     5221 gcatcatccc accacatgcc actctcgtct tcgatgtgga gcttctaaaa ctggaaggag
     5281 gaggaggtag tcctctgatt ctcaccctga gcctgattct ggtcgtgatt ctcgtcctcc
     5341 tcacagtcct ggccctcctg agccatcgga gagccctcaa acaaaaaatt tggcccggaa
     5401 ttccctcccc cgaatccgaa ttcgagggac tgtttacaac acataaaggc aattttcaac
     5461 tctggctcta tcaaaacgac ggatgtctct ggtggtcccc ttgtacacct tttaccgaag
     5521 atccccccgc cagcctcgag gtgctgagcg aacggtgttg gggcaccatg caagccgtcg
     5581 aacccggcac cgacgacgaa ggacctctcc tcgaacccgt cggaagcgaa cacgctcaag
     5641 acacatacct cgtcctcgat aagtggctgc tccctagaaa tccccctagc gaagatctgc
     5701 ccggccccgg cggaagcgtc gatatcgtcg ctatggacga gggaagcgag gccagcagct
     5761 gtagcagcgc cctggctagc aaaccttccc ccgaaggcgc tagcgccgct tccttcgaat
     5821 ataccattct cgatccttcc agccaactgc tgcggcccta actgtgcctt ctagttgcca
     5881 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     5941 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     6001 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     6061 tgctggggat gcggtgggct ctatggacac tgtgccctga gctgccccct accccacccc
     6121 acctaaagta
//
