LOCUS       sequence-366636-        5089 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 5089)
  TITLE     T4 Lysozyme Plasmids - Tray 4
REFERENCE   2  (bases 1 to 5089)
  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"
COMMENT     Sequence Label: T4 Lysozyme R119A Q123A WT*
FEATURES             Location/Qualifiers
     source          1..5089
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(13..18,19..34,35..41)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    49..65
                     /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)."
     promoter        join(108..113,114..129,130..136)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    144..160
                     /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(904..922)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        990..1094
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1095..1163,1164..1955)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1313..1332)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2126..2714
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2615..2634
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2868..2885
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2900..3040
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2966..2985)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3126..3148
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3161..3622)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVLPRAFR"
     primer_bind     3571..3593
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3585..3602
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3586..3602
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3618..3634)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3618..3634)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3642..3658
                     /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)."
     promoter        complement(join(3666..3672,3673..3690,3691..3696))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             complement(3745..4827)
                     /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     4789..4808
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4828..4905)
                     /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."
     promoter        join(4993..4998,4999..5016,5017..5023)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    5031..5047
                     /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)."
ORIGIN
        1 cccatccccc tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat
       61 aacaatttca cacaggaaac aggatcagct tactccccat ccccctgttg acaattaatc
      121 atcggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag gaaacaggat
      181 ccatcgatgc ttaggaggta ttatgaatat atttgaaatg ttacgtatag atgaaggtct
      241 tagacttaaa atctataaag acacagaagg ctattacact attggcatcg gtcatttgct
      301 tacaaaaagt ccatcactta atgctgctaa atctgaatta gataaagcta ttgggcgtaa
      361 tactaatggt gtaattacaa aagatgaggc tgaaaaactc tttaatcagg atgttgatgc
      421 tgctgttcgc ggaattctga gaaatgctaa attaaaaccg gtttatgatt ctcttgatgc
      481 ggttcgtcgc gctgcattga ttaatatggt tttccaaatg ggagaaaccg gtgtggcagg
      541 atttactaac tctttagcta tgcttcaagc aaaacgctgg gatgaagcag cagttaactt
      601 agctaaaagt agatggtata atcaaacacc taatcgcgca aaacgagtca ttacaacgtt
      661 tagaactggc acttgggacg cgtataaaaa tctataaagc tgtttacttt ctcttggaat
      721 tgtgatagta tattcacaat tacttgaata gacaattact aattaaaata tttaaaggaa
      781 acatatgaaa acatatcaag aatttattgc cgaagcttat cgatgataag ctgtcaaaca
      841 tgagcagatc tgagcccgcc taatgagcgg gctttttttt cagatctgct tgaagacgaa
      901 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
      961 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1021 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1081 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1141 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1201 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1261 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1321 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1381 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     1441 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1501 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     1561 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     1621 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     1681 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     1741 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     1801 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     1861 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     1921 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     1981 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2041 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2101 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2161 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2221 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     2281 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2341 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2401 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2461 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2521 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2581 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2641 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2701 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2761 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2821 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2881 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     2941 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     3001 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
     3061 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     3121 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     3181 gaacgccatc aaaaataatt cgcgtctggc cttcctgtag ccagctttca tcaacattaa
     3241 atgtgagcga gtaacaaccc gtcggattct ccgtgggaac aaacggcgga ttgaccgtaa
     3301 tgggataggt cacgttggtg tagatgggcg catcgtaacc gtgcatctgc cagtttgagg
     3361 ggacgacgac agtatcggcc tcaggaagat cgcactccag ccagctttcc ggcaccgctt
     3421 ctggtgccgg aaaccaggca aagcgccatt cgccattcag gctgcgcaac tgttgggaag
     3481 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     3541 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     3601 gtgaatccgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     3661 ccacacatta tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     3721 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3781 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     3841 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     3901 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     3961 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     4021 cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     4081 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     4141 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     4201 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     4261 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     4321 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     4381 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     4441 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     4501 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     4561 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     4621 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     4681 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     4741 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     4801 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     4861 ctatcatgcc ataccgcgaa aggttttgca ccattcgatg gtgtcctggc acgacaggtt
     4921 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta
     4981 ggcaccccag gctttacact ttatgcttcc ggctcgtata atgtgtggaa ttgtgagcgg
     5041 ataacaattt cacacaggaa acaggatcga tccatcgatg agcttactc
//
