LOCUS       sequence-382960-        5821 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  antibiotic resistance enzyme with tetracysteine tag for 
            visualization of enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5821)
  AUTHORS   Yeong V, Wang JW, Horn JM, Obermeyer AC
  TITLE     Intracellular phase separation of globular proteins facilitated by 
            short cationic peptides.
  JOURNAL   Nat Commun. 2022 Dec 22;13(1):7882. doi: 10.1038/s41467-022-35529-2.
  PUBMED    36550144
REFERENCE   2  (bases 1 to 5821)
  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: "Nat 
            Commun."; date: "2022-12-22"; volume: "13(1):7882. doi"; pages: " 
            10.1038/s41467-022-35529-2"
COMMENT     Sequence Label: pVY-14 TC-CAT
FEATURES             Location/Qualifiers
     source          1..5821
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     158..180
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(174..365)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(937..2019)
                     /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     1981..2000
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2020..2097)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2303..2322
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2410..2429
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2410..2428
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2429..2453
                     /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             2468..2490
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2502..2519
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     CDS             2529..3185
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="EKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFLK
                     TVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSSL
                     WSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANMD
                     NFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2592..2611)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3196..3240
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3278..3296)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3292..3339
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3376..3831
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3463..3482)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3673..3694
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(3946..4737,4738..4806))
                     /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     4569..4588
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4807..4899)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4980..5568
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5469..5488
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5722..5739
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(5754..5821,1..72)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(join(5819..5821,1..17))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
       61 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
      121 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
      181 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
      241 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
      301 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
      361 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
      421 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
      481 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
      541 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
      601 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
      661 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
      721 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
      781 cgccagccta gccgggtcct caacgacagg agcacgatca tgctagtcat gccccgcgcc
      841 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
      901 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
      961 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     1021 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     1081 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     1141 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     1201 gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     1261 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     1321 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     1381 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     1441 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     1501 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     1561 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     1621 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     1681 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     1741 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     1801 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     1861 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     1921 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     1981 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     2041 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     2101 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     2161 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     2221 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     2281 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     2341 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc
     2401 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
     2461 aaataatttt gtttaacttt aagaaggaga tatacatggg ttgttgccct gggtgttgtg
     2521 gtggcgctga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     2581 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     2641 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     2701 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag
     2761 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     2821 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     2881 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     2941 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     3001 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     3061 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
     3121 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     3181 cgtaataagt ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg
     3241 tctactagcg cagcttaatt aacctaggct gctgccaccg ctgagcaata actagcataa
     3301 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     3361 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3421 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3481 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3541 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3601 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3661 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3721 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3781 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt tctggcggca
     3841 cgatggcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3901 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3961 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4021 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4081 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4141 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4201 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4261 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4321 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4381 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4441 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4501 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4561 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4621 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4681 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4741 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4801 actcatactc ttcctttttc aatcatgatt gaagcattta tcagggttat tgtctcatga
     4861 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggtcatgac caaaatccct
     4921 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4981 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5041 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5101 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     5161 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5221 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5281 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttggt gcgaacgacc
     5341 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5401 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5461 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5521 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5581 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5641 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     5701 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     5761 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     5821 c
//
