LOCUS       sequence-315204-        4187 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NERD-C static protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4187)
  AUTHORS   Davey JA, Damry AM, Goto NK, Chica RA
  TITLE     Rational design of proteins that exchange on functional timescales.
  JOURNAL   Nat Chem Biol. 2017 Dec;13(12):1280-1285. doi: 
            10.1038/nchembio.2503. Epub 2017 Oct 23.
  PUBMED    29058725
REFERENCE   2  (bases 1 to 4187)
  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: 
            "10.1038/nchembio.2503"; journalName: "Nat Chem Biol"; date: 
            "2017-12"; volume: "13"; issue: "12"; pages: "1280-1285"
COMMENT     Sequence Label: pJ414 - NERD-C
FEATURES             Location/Qualifiers
     source          1..4187
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     858..875
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     859..875
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1338..1357
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1338..1356
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1357..1381
                     /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)."
     CDS             1427..1444
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1629..1676
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1899..1918)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1924..1943)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        1985..2076
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2077..2145,2146..2937)
                     /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(2295..2314)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             3020..4102
                     /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     complement(3039..3058)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag cccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag gcgagagtag
      781 ggaactgcca ggcatcaaac taagcagaag gcccctgacg gatggccttt ttgcgtttct
      841 acaaactctt tctgtgttgt aaaacgacgg ccagtcttaa gctcgggccc cctgggcggt
      901 tctgataacg agtaatcgtt aatccgcaaa taacgtaaaa acccgcttcg gcgggttttt
      961 ttatgggggg agtttaggga aagagcattt gtcagaatat ttaagggcgc ctgtcacttt
     1021 gcttgatata tgagaattat ttaaccttat aaatgagaaa aaagcaacgc actttaaata
     1081 agatacgttg ctttttcgat tgatgaacac ctataattaa actattcatc tattatttat
     1141 gattttttgt atatacaata tttctagttt gttaaagaga attaagaaaa taaatctcga
     1201 aaataataaa gggaaaatca gtttttgata tcaaaattat acatgtcaac gataatacaa
     1261 aatataatac aaactataag atgttatcag tatttattat catttagaat aaattttgtg
     1321 tcgcccttcc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc
     1381 ccctctagaa ataattttgt ttaactttta ggaggtaaaa catatgcacc accatcacca
     1441 ccatggtatg acgttcaagc tgattatcaa cggtaaaacg ctgaaaggcg agactaccac
     1501 cgaagccgtt gacgcggcta cggcagaaaa ggtcctgaaa cagtacgcga acgacaatgg
     1561 tatcgacggc gagtggacct atgatgatgc gaccaagacg tttaccgtga ccgagtaact
     1621 cgagccccct agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgcccc
     1681 tgagacgcgt caatcgagtt cgtacctaag ggcgacaccc cataattagc ccgggcgaaa
     1741 ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg
     1801 catggggagt ccccacacta ccatcggcgc tacggcgttt cacttctgag ttcggcatgg
     1861 ggtcaggtgg gaccaccgcg ctactgccgc caggcaaaca aggggtgtta tgagccatat
     1921 tcaggtataa atgggctcgc gataatgttc agaattggtt aattggttgt aacactgacc
     1981 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2041 tgataaatgc ttcaataata ttgaaaaagg aagaatatga gtattcaaca tttccgtgtc
     2101 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     2161 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2221 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2281 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     2341 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2401 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2461 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2521 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2581 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcgatggc aacaacgttg
     2641 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2701 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2761 attgctgata aatccggagc cggtgagcgt ggttctcgcg gtatcatcgc agcgctgggg
     2821 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2881 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaagcg
     2941 gcgcgccatc gaatggcgca aaacctttcg cggtatggca tgatagcgcc cggaagagag
     3001 tcaattcagg gtggtgaata tgaaaccagt aacgttatac gatgtcgcag agtatgccgg
     3061 tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac
     3121 gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca
     3181 acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca
     3241 cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt
     3301 ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct
     3361 tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat
     3421 tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac
     3481 acccatcaac agtattattt tctcccatga ggacggtacg cgactgggcg tggagcatct
     3541 ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc
     3601 gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc
     3661 ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa
     3721 tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat
     3781 gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga
     3841 cgataccgaa gatagctcat gttatatccc gccgttaacc accatcaaac aggattttcg
     3901 cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa
     3961 gggcaatcag ctgttgccag tctcactggt gaaaagaaaa accaccctgg cgcccaatac
     4021 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     4081 ccgactggaa agcgggcagt gactcatgac caaaatccct taacgtgagt tacgcgcgcg
     4141 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttc
//
