LOCUS       sequence-437013-        5600 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A plasmid encoding the hydrogel-forming protein PXP with the Q54A 
            coil mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5600)
  AUTHORS   Bennett JI, Boit MO, Gregorio NE, Zhang F, Kibler RD, Hoye JW, Prado
            O, Rapp PB, Murry CE, Stevens KR, DeForest CA
  TITLE     Genetically Encoded XTEN-based Hydrogels with Tunable 
            Viscoelasticity and Biodegradability for Injectable Cell Therapies.
  JOURNAL   Adv Sci (Weinh). 2024 Mar 13:e2301708. doi: 10.1002/advs.202301708.
  PUBMED    38477407
REFERENCE   2  (bases 1 to 5600)
  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: "Adv Sci 
            (Weinh). 2024 Mar 13:e2301708. doi: 10.1002/advs.202301708."
COMMENT     Sequence Label: Q54A
FEATURES             Location/Qualifiers
     source          1..5600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     170..189
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(408..512)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     580..598
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(622..636,637..642,643..659,660..666)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    642..658
                     /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)."
     protein_bind    674..690
                     /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)."
     RBS             709..720
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             739..756
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1630..1647
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1669..1763
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1807..2466
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1873..1892)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2531..2617
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2710..3792)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     3754..3773
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3793..3870)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(3954..3976)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4062..4202
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4117..4136
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4217..4234)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4388..4976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4468..4487)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5147..5600,1..338,339..407))
                     /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"
ORIGIN
        1 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
       61 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
      121 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
      181 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
      241 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
      301 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
      361 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
      421 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
      481 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
      541 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
      601 tttcgtcttc acgtcgagaa atcataaaaa atttatttgc tttgtgagcg gataacaatt
      661 ataatagatt caattgtgag cggataacaa tttcacacag aattcattaa agaggagaaa
      721 ttaactatga gaggatcgca tcaccatcac catcacggat ccggtcgtgg tgactctggc
      781 tcaggttcgg gtagtgggag cggctctggc agcggtgcgc cgcaaatgct gcgtgaactg
      841 caggaaacca atgccgcgct tcaggatgtg cgggaattgc ttcgtcaagc ggtcaaggag
      901 ataacgttct tgaagaacac cgtcatggag tcggatgcgt ctggctccgg aagcggaagt
      961 ggttctggta gcgtcgacgg ctccccggct ggcagcccga ccagcactga agagggcacg
     1021 agcgagtcgg cgaccccgga gtctggtccg ggcacctcca ccgaaccgtc tgagggcagc
     1081 gcaccgggta gcccggccgg tagccctacc agcaccgaag agggtaccag cacggaaccg
     1141 agcgaaggct cggcaccggg tacgagcacc gagccgtccg agggttccgc gccaggtacc
     1201 agcgagagcg caacgccgga gtccggtccg ggcagcgaac cagcgaccag cggcagcgaa
     1261 acgccgggtt cagagccggc gacgagcggt agcgagactc cgggcagccc ggctggtagc
     1321 ccgacgtcca ccgaagaagg caccagcgaa agcgccaccc cggagagcgg tcctggtacg
     1381 tctaccgagc catctgaggg tagcgcgccg ctcgagtcag gttcgggtag tgggagcggc
     1441 tctggcagcg gtgcgccgca aatgctgcgt gaactgcagg aaaccaatgc cgcgcttcag
     1501 gatgtgcggg aattgcttcg tcaagcggtc aaggagataa cgttcttgaa gaacaccgtc
     1561 atggagtcgg atgcgtccgg atctggctcc ggaagcggaa gtggttctgg tagcggtcgt
     1621 ggtgactctc atcaccatca ccatcacaag cttaattagc tgagcttgga ctcctgttga
     1681 tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc
     1741 gggcgttttt tattggtgag aatccaagct agcttggcga gattttcagg agctaaggaa
     1801 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
     1861 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
     1921 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     1981 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa
     2041 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     2101 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     2161 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     2221 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
     2281 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     2341 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     2401 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg
     2461 gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc ctggggtaat gactctctag
     2521 cttgaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     2581 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccctctagat tacgtgcagt
     2641 cgatgataag ctgtcaaaca tgagaattgt gcctaatgag tgagctaact tacattaatt
     2701 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     2761 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
     2821 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
     2881 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
     2941 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc
     3001 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
     3061 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
     3121 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
     3181 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
     3241 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
     3301 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
     3361 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
     3421 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
     3481 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
     3541 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
     3601 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
     3661 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     3721 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     3781 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
     3841 gcgaaaggtt ttgcaccatt cgatggtgtc ggaatttcgg gcagcgttgg gtcctggcca
     3901 cgggtgcgca tgatctagag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     3961 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     4021 gcccgtcagg gcgcgtcagc gggtgttggc gggggtcggg gcgcagccat gacccagtca
     4081 cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag attgtactga
     4141 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     4201 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4261 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4321 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4381 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4441 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4501 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4561 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4621 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4681 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4741 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4801 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     4861 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4921 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4981 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5041 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5101 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5161 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5221 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5281 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5341 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5401 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5461 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5521 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5581 gtcctccgat cgttgtcaga
//
