LOCUS       sequence-354765-        5393 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides the coding region to express the EcoRI restriction enzyme. 
            The RTX tag allows the EcoRI enzyme to be isolated via 
            calcium-precipitation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 114 to 148)
  AUTHORS   Hendrix J, Read T, Lalonde JF, Jensen PK, Heymann W, Lovelace E, 
            Zimmermann SA, Brasino M, Rokicki J, Dowell RD
  TITLE     Engineered calcium-precipitable restriction enzyme.
  JOURNAL   ACS Synth Biol. 2014 Dec 19;3(12):969-71. doi: 10.1021/sb500042m.
  PUBMED    25524101
REFERENCE   2  (bases 1 to 5393)
  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"; journalName: 
            "ACS Synth Biol"; date: "2014-12-19- 19"; volume: "3"; issue: "12"; 
            pages: "969-71"
COMMENT     Sequence Label: pSB1A3 EcoRI-RTX with EcoRI Methylase-AmilCP
FEATURES             Location/Qualifiers
     source          1..5393
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..42
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(46..89)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    92..113
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        114..148
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     RBS             157..168
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    3352..3372
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3373..3430
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(3454..3471)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3625..4213)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3705..3724)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4384..5175,5176..5244))
                     /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     5007..5026
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5245..5349)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttatcatgac attaacctat aaaaataggc gtatcacgag gcagaatttc agataaaaaa
       61 aatccttagc tttcgctaag gatgatttct ggaattcgcg gccgcttcta gagttgacgg
      121 ctagctcagt cctaggtaca gtgctagcta ctagagaaag aggagaaata ctagatggcc
      181 ggcgcacgca acgcgaccaa taagctcctg cacaaagcga aaaaaagcaa aagcgatgag
      241 ttttataccc agtattgcga cattgaaaac gaattacaat actaccgcga acatttcagc
      301 gataaggttg tgtactgcaa ctgtgatgac ccgcgtgtga gcaatttttt taaatatttc
      361 gcggtgaact ttgacaacct cggactgaag aagctgattg caagctgtta tgtagaaaac
      421 aaggagggct ttagctccag cgaagcagca aaaaatggct tttactatga ataccataaa
      481 gaaaacggca aaaaactggt cttcgatgat atttcggtga gtagcttctg tggcgacgga
      541 gattttcgga gcagtgaaag cattgatctg ctcaaaaaaa gtgatatcgt ggtcacgaac
      601 ccgccttttt cactctttcg tgaatactta gatcaactta ttaaatacga taaaaagttc
      661 cttatcatcg ccaatgtaaa cagcattact tataaagaag tgtttaatct gatcaaagaa
      721 aacaagatct ggctgggagt acatttaggc cggggtgtga gtggcttcat cgttccagaa
      781 cattacgaac tgtacggcac cgaggcccgc attgattcaa atggaaaccg tattatcagc
      841 ccgaataact gtctgtggct gacgaacctt gacgtcttca ttcgccacaa agacctgccg
      901 ttaacacgta aatactttgg aaacgaaagc tcgtacccaa agtatgataa ctatgatgcg
      961 attaacgtta acaagacgaa ggatattccg ctggattata atggcgttat gggggtgccg
     1021 atcaccttcc tgcataaatt caatcctgaa caatttgaat tgattaaatt ccggaaaggt
     1081 gtggatgaga aagacctatc aattaacgga aagtgcccat attttcgaat cctgattaaa
     1141 aataagcgct tacagaaaac cggcagtgtg atcgctaaac aaatgaccta caaggtttat
     1201 atgtcaggca cggtcaatgg acactacttt gaggtcgaag gcgatggaaa aggtaagccc
     1261 tacgaggggg agcagacggt aaagctcact gtcaccaagg gcggacctct gccatttgct
     1321 tgggatattt tatcaccaca gtgtcagtac ggaagcatac cattcaccaa gtaccctgaa
     1381 gacatccctg actatgtaaa gcagtcattc ccggagggct atacatggga gaggatcatg
     1441 aactttgaag atggtgcagt gtgtactgtc agcaatgatt ccagcatcca aggcaactgt
     1501 ttcatctacc atgtcaagtt ctctggtttg aactttcctc ccaatggacc tgtcatgcag
     1561 aagaagacac agggctggga acccaacact gagcgtctct ttgcacgaga tggaatgctg
     1621 ctaggaaaca actttatggc tctgaagtta gaaggaggcg gtcactattt gtgtgaattt
     1681 aaaactactt acaaggcaaa gaagcctgtg aagatgccag ggtatcacta tgttgaccgc
     1741 aaactggatg taaccaatca caacaaggat tacacttcgg ttgagcagtg tgaaatttcc
     1801 attgcacgca aacctgtggt cgccaccggt taatactaga tggccggcag caacaaaaaa
     1861 cagagcaatc gcctgactga gcagcacaaa ctgagccaag gtgttattgg catctttggc
     1921 gattatgcca aagcccacga cctggctgtg ggggaggtaa gcaaattggt taagaaggca
     1981 ctgagcaatg aatacccgca gctgtcgttc cgctatcgtg atagcatcaa aaaaacggaa
     2041 ataaatgaag ctctcaaaaa gattgatcca gacctgggtg gcactctttt tgtatcgaat
     2101 agctctatta agcctgatgg cgggattgtg gaagtgaagg atgattacgg ggagtggcgc
     2161 gtcgtgttag ttgccgaagc aaagcaccag ggtaaggata tcatcaacat tcgtaatggc
     2221 ttgcttgtcg gcaaacgtgg tgaccaggac ctgatggccg cgggcaacgc catcgagcgt
     2281 agccataaaa atatctcaga aatcgcaaat ttcatgttat cagaaagcca ttttccatat
     2341 gtgttatttc tggagggctc aaatttcttg accgaaaata tcagcattac ccgtccggat
     2401 ggtcgcgtgg ttaacctaga atataacagc ggcatcctga atcgtctgga tcgtctgacc
     2461 gcggctaact atggcatgcc aatcaactct aacctgtgca taaacaaatt cgtgaatcat
     2521 aaagataaat ctattatgct ccaggcggcg agcatttata cccaggggga tggtcgcgaa
     2581 tgggattcga aaatcatgtt cgagattatg tttgatatta gcactacgtc acttcgcgtg
     2641 ctgggtcgcg atctattcga gcagctgact agcaaaaccg gtatggccgg cggtgcgggc
     2701 aacgataccc tgtatggtgg cgccgggaat gacacattat acggaggtgc tggcaatgat
     2761 acgctgtatg gcggagcagg taacgacact ttgtatgggg gcgccggtaa cgacacgctg
     2821 tatggggggg caggcaacga taccctttat ggcggtgctg ggaatgacac actgtacggc
     2881 ggggcgggta acgataccct ctataccggc ggtgcgggca acgataccct gtatggtggc
     2941 gccgggaatg acacattata cggaggtgct ggcaatgata cgctgtatgg cggagcaggt
     3001 aacgacactt tgtatggggg cgccggtaac gacacgctgt atgggggggc aggcaacgat
     3061 accctttatg gcggtgctgg gaatgacaca ctgtacggcg gggcgggtaa cgataccctc
     3121 tataccggcg gtgcgggcaa cgataccctg tatggtggcg ccgggaatga cacattatac
     3181 ggaggtgctg gcaatgatac gctgtatggc ggagcaggta acgacacttt gtatgggggc
     3241 gccggtaacg acacgctgta tgggggggca ggcaacgata ccctttatgg cggtgctggg
     3301 aatgacacac tgtacggcgg ggcgggtaac gataccctct ataccggtta atactagtag
     3361 cggccgctgc agtccggcaa aaaagggcaa ggtgtcacca ccctgccctt tttctttaaa
     3421 accgaaaaga ttacttcgcg ttatgcaggc ttcctcgctc actgactcgc tgcgctcggt
     3481 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3541 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3601 taaaaaggcc gcgttgctgg cgtttttcca caggctccgc ccccctgacg agcatcacaa
     3661 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3721 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3781 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3841 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3901 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3961 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4021 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4081 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4141 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4201 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4261 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4321 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4381 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4441 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4501 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4561 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4621 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4681 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4741 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4801 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4861 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4921 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4981 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5041 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5101 atccagttcg atataaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5161 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5221 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5281 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5341 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca tta
//
