LOCUS       sequence-466145-        5943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV production plasmid for HBA1 UTRs vector from Fig. 3 that 
            mediates HDR at HBA1 locus using HBA1 sg5 gRNA. HBB gene includes 
            introns; HBA1 UTRs flank H+C15BA1 cassette. HAs are ~400bp each..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5943)
  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 5943)
  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: MKC175_HBA1reg(synEPOR)
FEATURES             Location/Qualifiers
     source          1..5943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     80..101
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(255..274)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     374..396
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(434..452)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        520..624
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(625..693,694..1485)
                     /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     complement(843..862)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1656..2244
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2145..2164
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2306..2435
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             3495..3815
                     /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    4401..4625
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5511..5651
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5511..5640
                     /label=AAV2 ITR
     rep_origin      join(5726..5943,1..238)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5813..5832)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
       61 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
      121 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
      181 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
      241 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
      301 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
      361 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
      421 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
      481 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
      541 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
      601 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
      661 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
      721 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
      781 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
      841 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
      901 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
      961 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     1021 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     1081 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     1141 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     1201 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     1261 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     1321 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     1381 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     1441 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     1501 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1561 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     1621 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1681 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1741 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1801 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1861 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1921 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     1981 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2041 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2101 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2161 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2221 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     2281 ttttgctggc cttttgctca catgtcctgc aggcagctgc gcgctcgctc gctcactgag
     2341 gccgcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag
     2401 agagggagtg gccaactcca tcactagggg ttcctgcgct ccagccggtt ccagctattg
     2461 ctttgtttac ctgtttaacc agtatttacc tagcaagtct tccatcagat agcatttgga
     2521 gagctggggg tgtcacagtg aaccacgacc tctaggccag tgggagagtc agtcacacaa
     2581 actgtgagtc catgacttgg ggcttagccc gcacccacca ccccacgcgc caccccacaa
     2641 ccccgggtag aggagtctga atctggagcc gccctcagcc cagccccctg ctttttgcgt
     2701 cctggtgttt attccttccc ggtgcccgtc actcaagcac actggtgact atcgccagag
     2761 ggaaagggag ctgcaggaag cgaggctgga gagcaggagg ggctctgcgc agaaattctt
     2821 ttgagttcct atgggccagg gcgtccgggt gcgcgcattc ctctccgccc caggattggg
     2881 cgaagcctcc cggctcgcac tcgctcgccc gtgtgttccc cgatcccgct ggagtcgatg
     2941 cgcgtccagc gcgtgccagg ccggggcggg ggtgggggct gcctttcccc cccgctgggg
     3001 ccgccccggc gccggaaagg aagggggggc gctgcgcccg ggggtgcacg agccgacagc
     3061 gcccgccccc aacgggccgg ccccgccagc gccgctaccg ccctgccccc gggcgagcgg
     3121 gatgggcggg agtggagtgg cgggtggagg gtggagacgt cctggccccc gccccgcgtg
     3181 cacccccagg ggaggccgag cccgccgccc ggccccgcgc aggccccgcc cgggactccc
     3241 ctgcggtcca ggccgcgccc cgggctccgc gccagccaat gagcgccgcc cggccgggcg
     3301 tgcccccgcg ccccaagcat aaaccctggc gcgctcgcgg cccggcactc ttctggtccc
     3361 cacagactca gagagaaccc accatgaact ccttctccac aagcgccttc ggtccagttg
     3421 ccttctccct ggggctgctc ctggtgttgc ctgctgcctt ccctgcccca ggcggcggcg
     3481 gaagcatgct tgagggagtg caggtggaaa ccatctcccc aggagacggg cgcaccttcc
     3541 ccaagcgcgg ccagacctgc gtggtgcact acaccgggat gcttgaagat ggaaagaaag
     3601 ttgattcctc ccgggacaga aacaagccct ttaagtttat gctaggcaag caggaggtga
     3661 tccgaggctg ggaagaaggg gttgcccaga tgagtgtggg tcagagagcc aaactgacta
     3721 tatctccaga ttatgcctat ggtgccactg ggcacccagg catcatccca ccacatgcca
     3781 ctctcgtctt cgatgtggag cttctaaaac tggaaggagg aggaggtagt cccctcatcc
     3841 tgacgctctc cctcatcctc gtggtcatcc tggtgctgct gaccgtgctc gcgctgctct
     3901 cccaccgccg ggctctgaag cagaagatct ggcctggcat cccgagccca gagagcgagt
     3961 ttgaaggcct cttcaccacc cacaagggta acttccagct gtggctgtac cagaatgatg
     4021 gctgcctgtg gtggagcccc tgcaccccct tcacggagga cccacctgct tccctggaag
     4081 tcctctcaga gcgctgctgg gggacgatgc aggcagtgga gccggggaca gatgatgagg
     4141 gccccctgct ggagccagtg ggcagtgagc atgcccagga tacctatctg gtgctggaca
     4201 aatggttgct gccccggaac ccgcccagtg aggacctccc agggcctggt ggcagtgtgg
     4261 acatagtggc catggatgaa ggctcagaag catcctcctg ctcatctgct ttggcctcga
     4321 agcccagccc agagggagcc tctgctgcca gctttgagta cactatcctg gaccccagct
     4381 cccagctctt gcgtccataa ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     4441 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     4501 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     4561 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     4621 tatggtggcc atgcttcttg ccccttgggc ctccccccag cccctcctcc ccttcctgca
     4681 cccgtacccc cgtggtcttt gaataaagtc tgagtgggcg gcagcctgtg tgtgcctgag
     4741 ttttttccct cagcaaacgt gccaggcatg ggtgtggaca gcagctggga cacacatggc
     4801 tagaacctct ctgcagctgg atagggtagg aaaaggcagg ggcgggagga ggggatggag
     4861 gagggaaagt ggagccaccg cgaagtccag ctggaaaaac gctggaccct agagtgcttt
     4921 gaggatgcat ttgctctttc ccgagtttta ttcccagact tttcagattc aatgcaggtt
     4981 tgctgaaata atgaatttat ccatctttac gtttctgggc actcttgtgc caagaactgg
     5041 ctggctttct gcctgggacg tcactggttt cccagaggtc ctcccacata tgggtggtgg
     5101 gtaggtcaga gaagtcccac tccagcatgg ctgcattgat cccccatcgt tcccactagt
     5161 ctccgtaaaa cctcccagat acaggcacag tctagatgaa atcaggggtg cggggtgcaa
     5221 ctgcaggccc caggcaattc aataggggct ctactttcac ccccaggtca ccccagaatg
     5281 ctcacacacc agacactgac gccctggggc tgtcaagatc aggcgtttgt ctctgggccc
     5341 agctcagggc ccagctcagc acccactcag ctcccctgag gctggggagc ctgtcccatt
     5401 gcgactggag aggagagcgg ggccacagag gcctggctag aaggtccctt ctccctggtg
     5461 tgtgttttct ctctgctgag caggcttgca gtgcctgggg tacaggccgc aggaacccct
     5521 agtgatggag ttggccactc cttctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     5581 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     5641 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     5701 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5761 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     5821 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     5881 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     5941 ttg
//
