LOCUS       sequence-443889-        5621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A plasmid encoding the hydrogel-forming protein PXP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5621)
  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 5621)
  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: PXP
FEATURES             Location/Qualifiers
     source          1..5621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     164..182
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(206..220,221..226,227..243,244..250)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    226..242
                     /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    258..274
                     /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             293..304
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             323..340
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1235..1252
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1274..1368
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1412..2071
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1478..1497)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2136..2222
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2315..3397)
                     /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     3359..3378
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3398..3475)
                     /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(3559..3581)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3667..3807
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3722..3741
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3822..3839)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3993..4581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4073..4092)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4752..5543,5544..5612))
                     /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     5375..5394
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5613..5621,1..96))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
       61 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
      121 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
      181 gccctttcgt cttcacgtcg agaaatcata aaaaatttat ttgctttgtg agcggataac
      241 aattataata gattcaattg tgagcggata acaatttcac acagaattca ttaaagagga
      301 gaaattaact atgagaggat cgcatcacca tcaccatcac ggatccggtc gtggtgactc
      361 tgtcgacggc tcaggttcgg gtagtgggag cggctctggc agcggtgcgc cgcaaatgct
      421 gcgtgaactg caggaaacca atgccgcgct tcaggatgtg cgggaattgc ttcgtcaaca
      481 ggtcaaggag ataacgttct tgaagaacac cgtcatggag tcggatgcgt ccggatctgg
      541 ctccggaagc ggaagtggtt ctggtagcct cgacggctcc ccggctggca gcccgaccag
      601 cactgaagag ggcacgagcg agtcggcgac cccggagtct ggtccgggca cctccaccga
      661 accgtctgag ggcagcgcac cgggtagccc ggccggtagc cctaccagca ccgaagaggg
      721 taccagcacg gaaccgagcg aaggctcggc accgggtacg agcaccgagc cgtccgaggg
      781 ttccgcgcca ggtaccagcg agagcgcaac gccggagtcc ggtccgggca gcgaaccagc
      841 gaccagcggc agcgaaacgc cgggttcaga gccggcgacg agcggtagcg agactccggg
      901 cagcccggct ggtagcccga cgtccaccga agaaggcacc agcgaaagcg ccaccccgga
      961 gagcggtcct ggtacgtcta ccgagccatc tgagggtagc gcgccgctcg acggctcagg
     1021 ttcgggtagt gggagcggct ctggcagcgg tgcgccgcaa atgctgcgtg aactgcagga
     1081 aaccaatgcc gcgcttcagg atgtgcggga attgcttcgt caacaggtca aggagataac
     1141 gttcttgaag aacaccgtca tggagtcgga tgcgtccgga tctggctccg gaagcggaag
     1201 tggttctggt agcctcgagg gtcgtggtga ctctcatcac catcaccatc acaagcttaa
     1261 ttagctgagc ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat
     1321 ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagctagctt
     1381 ggcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac
     1441 cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca
     1501 atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa
     1561 aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca
     1621 tccggaattt cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc
     1681 ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca
     1741 cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa
     1801 cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg
     1861 ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt
     1921 tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca
     1981 ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca
     2041 gtactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta
     2101 aacgcctggg gtaatgactc tctagcttga ggcatcaaat aaaacgaaag gctcagtcga
     2161 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     2221 atccgccctc tagattacgt gcagtcgatg ataagctgtc aaacatgaga attgtgccta
     2281 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2341 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2401 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     2461 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     2521 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     2581 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     2641 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     2701 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     2761 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     2821 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     2881 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     2941 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     3001 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     3061 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     3121 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     3181 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     3241 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     3301 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     3361 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     3421 cttccgggcg ctatcatgcc ataccgcgaa aggttttgca ccattcgatg gtgtcggaat
     3481 ttcgggcagc gttgggtcct ggccacgggt gcgcatgatc tagagctgcc tcgcgcgttt
     3541 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     3601 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     3661 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat
     3721 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     3781 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     3841 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3901 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3961 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4021 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4081 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4141 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4201 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4261 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4321 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4381 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     4441 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4501 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4561 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4621 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4681 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4741 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4801 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4861 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4921 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4981 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5041 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5101 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5161 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5221 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5281 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5341 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5401 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5461 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5521 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5581 ataagggcga cacggaaatg ttgaatactc atactcttcc t
//
