LOCUS       sequence-455995-        7288 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression in BL21(DE3) of T4 UvsY-like recombination mediator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7288)
  TITLE     Escherichia phage T4 Expression Constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7288)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pTBY1-T4uvsY
FEATURES             Location/Qualifiers
     source          1..7288
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..77
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      275..863
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     764..783
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1017..1034
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1056..1075)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1216..1238
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1232..1423)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1980..3062)
                     /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     3024..3043
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3063..3140)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      3293..3379
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3476..3562
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3697..3716
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3697..3715
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3716..3740
                     /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             3755..3777
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4196..5014
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae VMA1"
                     /product="5' region of the intein from the yeast Vma1 
                     subunit of the vacuolar ATPase (Chong et al., 1998)"
                     /label=Sce VMA intein 5' region
                     /translation="CFAKGTNVLMADGSIECIENIEVGNKVMGKDGRPREVIKLPRGRE
                     TMYSVVQKSQHRAHKSDSSREVPELLKFTCNATHELVVRTPRSVRRLSRTIKGVEYFEV
                     ITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELVESYRKASNKAYFEWTIEARD
                     LSLLGSHVRKATYQTYAPILYENDHFFDYMQKSKFHLTIEGPKVLAYLLGLWIGDGLSD
                     RATFSVDSRDTSLMERVTEYAEKLNLCAEYKDRKEPQVAKTVNLYSKVVRG"
     CDS             5470..5481
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             5591..5746
                     /codon_start=1
                     /gene="Bacillus circulans chiA"
                     /product="chitin binding domain from chitinase A1 (Watanabe
                     et al., 1994)"
                     /label=CBD
                     /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV
                     PALWQLQ"
     primer_bind     complement(5811..5829)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5825..5872
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        5932..6036
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6037..6105,6106..6897)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6255..6274)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(join(6939..7288,1..164))
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7134..7155)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
       61 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
      121 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtaaaagg atctaggtga
      181 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
      241 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
      301 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
      361 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
      421 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
      481 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
      541 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
      601 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
      661 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
      721 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
      781 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
      841 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
      901 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
      961 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     1021 agtcagtgag cgaggaagct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     1081 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
     1141 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     1201 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     1261 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg cagcgattca cagatgtctg
     1321 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     1381 taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag
     1441 ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac
     1501 gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg
     1561 tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagccg aacgccagca
     1621 agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac
     1681 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg
     1741 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc
     1801 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
     1861 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     1921 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     1981 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     2041 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     2101 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     2161 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     2221 catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc
     2281 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     2341 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     2401 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     2461 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     2521 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     2581 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     2641 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     2701 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     2761 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     2821 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     2881 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     2941 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     3001 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     3061 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     3121 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     3181 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg
     3241 ccggcatgcc gccctttcgt cttcaagaat taattcccaa ttccccaggc atcaaataaa
     3301 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     3361 tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg
     3421 agggtggcgg gcaggacgcc cgccataaac tgccaggaat taattcccca ggcatcaaat
     3481 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     3541 cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc
     3601 cggagggtgg cgggcaggac gcccgccata aactgccagg aattggggat cggaattaat
     3661 tcccggttta aaccggggat ctcgatcccg cgaaattaat acgactcact ataggggaat
     3721 tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat
     3781 acatatgaga ttagaagatc ttcaagaaga attgaagaaa gatgtgttta tagattcgac
     3841 taaattacag tatgaagcag ctaataatgt gatgttatat agtaaatggc ttaataagca
     3901 ttcaagtatt aaaaaggaaa tgcttagaat tgaagcacag aaaaaagttg ctcttaaagc
     3961 tagattagac tactactcgg gacgaggaga tggtgatgaa tttagtatgg atcgttacga
     4021 gaaatcagaa atgaagacag ttctatcagc ggataaggat gttttaaagg ttgatacctc
     4081 gttgcagtat tgggggattt tattagattt ctgtagcgga gctcttgatg ctattaaatc
     4141 acgtggattt gctattaagc atattcaaga catgcgagca tttgaggctg gaaaatgctt
     4201 tgccaagggt accaatgttt taatggcgga tgggtctatt gaatgtattg aaaacattga
     4261 ggttggtaat aaggtcatgg gtaaagatgg cagacctcgt gaggtaatta aattgcccag
     4321 aggaagagaa actatgtaca gcgtcgtgca gaaaagtcag cacagagccc acaaaagtga
     4381 ctcaagtcgt gaagtgccag aattactcaa gtttacgtgt aatgcgaccc atgagttggt
     4441 tgttagaaca cctcgtagtg tccgccgttt gtctcgtacc attaagggtg tcgaatattt
     4501 tgaagttatt acttttgaga tgggccaaaa gaaagccccc gacggtagaa ttgttgagct
     4561 tgtcaaggaa gtttcaaaga gctacccaat atctgagggg cctgagagag ccaacgaatt
     4621 agtagaatcc tatagaaagg cttcaaataa agcttatttt gagtggacta ttgaggccag
     4681 agatctttct ctgttgggtt cccatgttcg taaagctacc taccagactt acgctccaat
     4741 tctttatgag aatgaccact ttttcgacta catgcaaaaa agtaagtttc atctcaccat
     4801 tgaaggtcca aaagtacttg cttatttact tggtttatgg attggtgatg gattgtctga
     4861 cagggcaact ttttcggttg attccagaga tacttctttg atggaacgtg ttactgaata
     4921 tgctgaaaag ttgaatttgt gcgccgagta taaggacaga aaagaaccac aagttgccaa
     4981 aactgttaat ttgtactcta aagttgtcag aggtaatggt attcgcaata atcttaatac
     5041 tgagaatcca ttatgggacg ctattgttgg cttaggattc ttgaaggacg gtgtcaaaaa
     5101 tattccttct ttcttgtcta cggacaatat cggtactcgt gaaacatttc ttgctggtct
     5161 aattgattct gatggctatg ttactgatga gcatggtatt aaagcaacaa taaagacaat
     5221 tcatacttct gtcagagatg gtttggtttc ccttgctcgt tctttaggct tagtagtctc
     5281 ggttaacgca gaacctgcta aggttgacat gaatgtcacc aaacataaaa ttagttatgc
     5341 tatttatatg tctggtggag atgttttgct taacgttctt tcgaagtgtg ccggctctaa
     5401 aaaattcagg cctgctcccg ccgctgcttt tgcacgtgag tgccgcggat tttatttcga
     5461 gttacaagaa ttgaaggaag acgattatta tgggattact ttatctgatg attctgatca
     5521 tcagtttttg cttggatccc aggttgtcgt ccatgcatgc ggtggcctga ccggtctgaa
     5581 ctcaggcctc acgacaaatc ctggtgtatc cgcttggcag gtcaacacag cttatactgc
     5641 gggacaattg gtcacatata acggcaagac gtataaatgt ttgcagcccc acacctcctt
     5701 ggcaggatgg gaaccatcca acgttcctgc cttgtggcag cttcaatgac tgcaggaagg
     5761 ggatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     5821 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     5881 aggaactata tccggataac tacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5941 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     6001 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     6061 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6121 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6181 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgtttccc aatgatgagc
     6241 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     6301 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6361 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6421 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6481 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6541 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6601 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6661 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6721 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6781 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6841 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6901 tcagaccaag tttactcata tatactttag attgatttac cccggttgat aatcagaaaa
     6961 gccccaaaaa caggaagatt gtataagcaa atatttaaat tgtaaacgtt aatattttgt
     7021 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     7081 gcaaaatccc ttataaatca aaagaatagc ccgagatagg gttgagtgtt gttccagttt
     7141 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     7201 atcagggcga tggcccacta cgtgaaccat cacccaaatc aagttttttg gggtcgaggt
     7261 gccgtaaagc actaaatcgg aaccctaa
//
