LOCUS       sequence-455996-        7723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression in BL21(DE3) of T4 endonuclease V N-glycosylase UV repair
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7723)
  TITLE     Escherichia phage T4 Expression Constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7723)
  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            "Unpublished"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: pTYB1-T4denV
FEATURES             Location/Qualifiers
     source          1..7723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(27..113)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        266..343
                     /gene="lacI"
                     /label=lacI promoter
     CDS             344..1426
                     /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(363..382)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             1983..2174
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2168..2190)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2331..2350
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2372..2389)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2543..3131)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2623..2642)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      3242..3755
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3329..3348)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3539..3560
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(3797..4588,4589..4657))
                     /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     4420..4439
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4658..4762)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(4822..4869)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4865..4883
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4948..5103)
                     /codon_start=1
                     /gene="Bacillus circulans chiA"
                     /product="chitin binding domain from chitinase A1 (Watanabe
                     et al., 1994)"
                     /label=CBD
                     /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV
                     PALWQLQ"
     CDS             complement(5213..5224)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(5680..6498)
                     /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"
     gap             7050..7149
                     /estimated_length=100
     RBS             7281..7303
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    7318..7342
                     /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)."
     primer_bind     complement(7342..7361)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7343..7361)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(7496..7582)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
       61 caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctg
      121 gggaattggg aattaattct tgaagacgaa agggcggcat gccggcatgc accattcctt
      181 gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg
      241 cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct ttcgcggtat
      301 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
      361 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
      421 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
      481 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
      541 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
      601 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
      661 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
      721 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
      781 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
      841 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
      901 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
      961 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
     1021 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
     1081 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
     1141 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt
     1201 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
     1261 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
     1321 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     1381 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     1441 atgtaagtta gctcactcat taggcaccgg gatctcgacc gatgcccttg agagccttca
     1501 acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg
     1561 tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg
     1621 aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct
     1681 tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc
     1741 aggccattat cgccggcatg gcggccgacg cgctgggcta cgtcttgctg gcgttcggct
     1801 ggcattgacc ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac
     1861 cctcacaacg ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc
     1921 tctctcgttt catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat
     1981 cagtgaccaa acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat
     2041 taacgcttct ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat
     2101 cgctgcacga ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg
     2161 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     2221 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
     2281 ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga
     2341 gcagattgta ctgagagtgc accatagctt cctcgctcac tgactcgctg cgctcggtcg
     2401 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2461 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2521 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2581 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2641 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2701 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2761 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2821 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2881 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2941 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     3001 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3061 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3121 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3181 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3241 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3301 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3361 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3421 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     3481 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3541 ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta
     3601 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3661 aacgcgaatt ttaacaaaat attaacgttt acaatttaaa tatttgctta tacaatcttc
     3721 ctgtttttgg ggcttttctg attatcaacc ggggtaaatc aatctaaagt atatatgagt
     3781 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3841 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3901 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3961 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4021 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4081 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4141 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4201 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4261 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4321 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4381 tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca
     4441 gaactttaaa agtgctcatc attgggaaac gttcttcggg gcgaaaactc tcaaggatct
     4501 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4561 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4621 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4681 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4741 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtagttatcc
     4801 ggatatagtt cctcctttca gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg
     4861 ttatgctagt tattgctcag cggtggcagc agccaactca gcttcctttc gggctttgtt
     4921 agcagccgga tccccttcct gcagtcattg aagctgccac aaggcaggaa cgttggatgg
     4981 ttcccatcct gccaaggagg tgtggggctg caaacattta tacgtcttgc cgttatatgt
     5041 gaccaattgt cccgcagtat aagctgtgtt gacctgccaa gcggatacac caggatttgt
     5101 cgtgaggcct gagttcagac cggtcaggcc accgcatgca tggacgacaa cctgggatcc
     5161 aagcaaaaac tgatgatcag aatcatcaga taaagtaatc ccataataat cgtcttcctt
     5221 caattcttgt aactcgaaat aaaatccgcg gcactcacgt gcaaaagcag cggcgggagc
     5281 aggcctgaat tttttagagc cggcacactt cgaaagaacg ttaagcaaaa catctccacc
     5341 agacatataa atagcataac taattttatg tttggtgaca ttcatgtcaa ccttagcagg
     5401 ttctgcgtta accgagacta ctaagcctaa agaacgagca agggaaacca aaccatctct
     5461 gacagaagta tgaattgtct ttattgttgc tttaatacca tgctcatcag taacatagcc
     5521 atcagaatca attagaccag caagaaatgt ttcacgagta ccgatattgt ccgtagacaa
     5581 gaaagaagga atatttttga caccgtcctt caagaatcct aagccaacaa tagcgtccca
     5641 taatggattc tcagtattaa gattattgcg aataccatta cctctgacaa ctttagagta
     5701 caaattaaca gttttggcaa cttgtggttc ttttctgtcc ttatactcgg cgcacaaatt
     5761 caacttttca gcatattcag taacacgttc catcaaagaa gtatctctgg aatcaaccga
     5821 aaaagttgcc ctgtcagaca atccatcacc aatccataaa ccaagtaaat aagcaagtac
     5881 ttttggacct tcaatggtga gatgaaactt acttttttgc atgtagtcga aaaagtggtc
     5941 attctcataa agaattggag cgtaagtctg gtaggtagct ttacgaacat gggaacccaa
     6001 cagagaaaga tctctggcct caatagtcca ctcaaaataa gctttatttg aagcctttct
     6061 ataggattct actaattcgt tggctctctc aggcccctca gatattgggt agctctttga
     6121 aacttccttg acaagctcaa caattctacc gtcgggggct ttcttttggc ccatctcaaa
     6181 agtaataact tcaaaatatt cgacaccctt aatggtacga gacaaacggc ggacactacg
     6241 aggtgttcta acaaccaact catgggtcgc attacacgta aacttgagta attctggcac
     6301 ttcacgactt gagtcacttt tgtgggctct gtgctgactt ttctgcacga cgctgtacat
     6361 agtttctctt cctctgggca atttaattac ctcacgaggt ctgccatctt tacccatgac
     6421 cttattacca acctcaatgt tttcaataca ttcaatagac ccatccgcca ttaaaacatt
     6481 ggtacccttg gcaaagcatg cataaatcgc cttaccgtag tatttgtacc aagtaggacg
     6541 ttgtgcaatt ttttcatcta aacgagcttg tgatatagca atagaagctt catggggaat
     6601 ataatcacca cggaattcct gaggaatatc actaatatcc tggactgtag tatccttgat
     6661 attaaaacca cgttttaaac attcagctat aagctcaatt tgacgtttac gtaagaactc
     6721 gagcttatcg taaaagaatg taacatgacc tgcgccaagg ataaaagtag gactgatttt
     6781 aaaatcacga acacgtttac cgttagcaac atgcttacga actgcaccaa aaacacgcgg
     6841 caattcacga tattcagcca ttaagtgttg gtcagccaat tcagatacta aagtaaggtt
     6901 gatacgagtc atatgctctt ccgcatgcat aaatcgcctt accgtagtat ttgtaccaag
     6961 taggacgttg tgcaattttt tcatctaaac gagcttgtga tatagcaata gaagcttcat
     7021 ggggaatata atcaccacgg aattcctgan nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     7081 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     7141 nnnnnnnnng aacacgttta ccgttagcaa catgcttacg aactgcacca aaaacacgcg
     7201 gcaattcacg atattcagcc attaagtgtt ggtcagccaa ttcagatact aaagtaaggt
     7261 tgatacgagt catatgtata tctccttctt aaagttaaac aaaattattt ctagagggga
     7321 attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg gatcgagatc
     7381 cccggtttaa accgggaatt aattccgatc cccaattcct ggcagtttat ggcgggcgtc
     7441 ctgcccgcca ccctccgggc cgttgcttcg caacgttcaa atccgctccc ggcggatttg
     7501 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtctttcg
     7561 actgagcctt tcgttttatt tgatgcctgg ggaattaatt cctggcagtt tatggcgggc
     7621 gtcctgcccg ccaccctccg ggccgttgct tcgcaacgtt caaatccgct cccggcggat
     7681 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaa
//
