LOCUS       sequence-42021-e        5887 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5887)
  AUTHORS   Uttamapinant C, White KA, Baruah H, Thompson S, Fernandez-Suarez M, 
            Puthenveetil S, Ting AY
  TITLE     A fluorophore ligase for site-specific protein labeling inside 
            living cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2010 Jun 15. 107(24):10914-9.
  PUBMED    20534555
REFERENCE   2  (bases 1 to 5887)
  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: "Proc Natl 
            Acad Sci U S A. 2010 Jun 15. 107(24):10914-9."
COMMENT     Sequence Label: pYFJ16-LplA(W37V) (for E. Coli expression)
FEATURES             Location/Qualifiers
     source          1..5887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..24,25..30,31..47,48..54)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /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    62..78
                     /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             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             121..141
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     terminator      1344..1438
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1482..2141
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1548..1567)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2206..2292
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2385..3467)
                     /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     3429..3448
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3468..3545)
                     /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(3629..3651)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3737..3877
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3792..3811
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3892..3909)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4063..4651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4143..4162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4822..5613,5614..5682))
                     /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     5445..5464
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5683..5787)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5855..5873
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaaa
      121 catcaccatc accatcacca tatgtccaca ttacgcctgc tcatctctga ctcttacgac
      181 ccgtggttta acctggcggt ggaagagtgt atttttcgcc aaatgcccgc cacgcagcgc
      241 gttctgtttc tcgtgcgcaa tgccgacacg gtagtaattg gtcgcgcgca gaacccgtgg
      301 aaagagtgta atacccggcg gatggaagaa gataacgtcc gcctggcgcg gcgcagtagc
      361 ggtggcggcg cggtgttcca cgatctcggc aatacctgct ttacctttat ggctggcaag
      421 ccggagtacg ataaaactat ctccacgtcg attgtgctca atgcgctgaa cgcgctcggc
      481 gtcagcgccg aagcgtccgg acgtaacgat ctggtggtga aaaccgtcga aggcgaccgc
      541 aaagtctcag gctcggccta tcgcgaaacc aaagatcgcg gcttccacca cggcaccttg
      601 ctactcaatg ccgacctcag ccgcctggca aactatctca atccggataa aaagaaactg
      661 gcggcgaaag gcattacgtc ggtacgttcc cgcgtgacca acctcaccga gctgttgccg
      721 gggatcaccc atgagcaggt ttgcgaggcc ataaccgagg cctttttcgc ccattatggc
      781 gagcgcgtgg aagcggaaat catctccccg aacaaaacgc cagacttgcc aaacttcgcc
      841 gaaacctttg cccgccagag tagctgggaa tggaacttcg gtcaggctcc ggcattctcg
      901 catctgctgg atgaacgctt tacctggggc ggcgtggaac tgcatttcga cgttgaaaaa
      961 ggccatatca cccgcgccca ggtgtttacc gacagcctca accccgcgcc gctggaagcc
     1021 ctcgccggac gactgcaagg ctgcctgtac cgcgcagata tgctgcaaca ggagtgcgaa
     1081 gcgctgttgg ttgacttccc ggaacaggaa aaagagctac gggagttatc ggcatggatg
     1141 gcgggggctg taaggtagtt acccgcccat gcgggcaact ttctcttcga tttgccggat
     1201 tttttccggc atttcttgcg caatgtggcg aaacgcctcg ctgatacgcg gatctctggc
     1261 gaccgtttgc cagaactgct gcgccagcgg gatcgccgtt ggggatccgt cgacctgcag
     1321 ccaagcttaa ttagctgagc ttggactcct gttgatagat ccagtaatga cctcagaact
     1381 ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc
     1441 aagctagctt ggcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg
     1501 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
     1561 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga
     1621 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
     1681 tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg atatgggata
     1741 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga
     1801 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt
     1861 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag
     1921 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct
     1981 tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc
     2041 tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg
     2101 aattacaaca gtactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt
     2161 ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat aaaacgaaag
     2221 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     2281 agtaggacaa atccgccctc tagattacgt gcagtcgatg ataagctgtc aaacatgaga
     2341 attgtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc
     2401 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2461 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
     2521 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
     2581 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
     2641 gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg
     2701 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
     2761 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
     2821 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
     2881 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
     2941 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
     3001 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
     3061 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
     3121 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
     3181 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
     3241 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
     3301 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
     3361 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
     3421 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     3481 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgca ccattcgatg
     3541 gtgtcggaat ttcgggcagc gttgggtcct ggccacgggt gcgcatgatc tagagctgcc
     3601 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     3661 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     3721 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
     3781 gcttaactat gcggcatcag agcagattgt actgagagtg caccacatgc ggtgtgaaat
     3841 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
     3901 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3961 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4021 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4081 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4141 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4201 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4261 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4321 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4381 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4441 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4501 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4561 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4621 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4681 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4741 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4801 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4861 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4921 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4981 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5041 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5101 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5161 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5221 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5281 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5341 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5401 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5461 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5521 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5581 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5641 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5701 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5761 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5821 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5881 tcttcac
//
