LOCUS       sequence-202584-        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 3
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 L99A WT*
FEATURES             Location/Qualifiers
     source          1..5089
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(728..746)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        814..918
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(919..987,988..1779)
                     /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(1137..1156)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1950..2538
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2439..2458
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2692..2709
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2724..2864
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2790..2809)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2950..2972
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2985..3446)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVLPRAFR"
     primer_bind     3395..3417
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3409..3426
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3410..3426
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3442..3458)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3442..3458)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3466..3482
                     /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(3490..3496,3497..3514,3515..3520))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             complement(3569..4651)
                     /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     4613..4632
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4652..4729)
                     /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(4817..4822,4823..4840,4841..4847)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    4855..4871
                     /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(4926..4931,4932..4947,4948..4954)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4962..4978
                     /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(5021..5026,5027..5042,5043..5049)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    5057..5073
                     /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 ggatccatcg atgcttagga ggtattatga atatatttga aatgttacgt atagatgaag
       61 gtcttagact taaaatctat aaagacacag aaggctatta cactattggc atcggtcatt
      121 tgcttacaaa aagtccatca cttaatgctg ctaaatctga attagataaa gctattgggc
      181 gtaatactaa tggtgtaatt acaaaagatg aggctgaaaa actctttaat caggatgttg
      241 atgctgctgt tcgcggaatt ctgagaaatg ctaaattaaa accggtttat gattctcttg
      301 atgcggttcg tcgcgctgca gcgattaata tggttttcca aatgggagaa accggtgtgg
      361 caggatttac taactcttta cgtatgcttc aacaaaaacg ctgggatgaa gcagcagtta
      421 acttagctaa aagtagatgg tataatcaaa cacctaatcg cgcaaaacga gtcattacaa
      481 cgtttagaac tggcacttgg gacgcgtata aaaatctata aagctgttta ctttctcttg
      541 gaattgtgat agtatattca caattacttg aatagacaat tactaattaa aatatttaaa
      601 ggaaacatat gaaaacatat caagaattta ttgccgaagc ttatcgatga taagctgtca
      661 aacatgagca gatctgagcc cgcctaatga gcgggctttt ttttcagatc tgcttgaaga
      721 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
      781 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
      841 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
      901 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
      961 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1021 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1081 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1141 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     1201 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1261 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1321 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     1381 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     1441 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     1501 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     1561 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     1621 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     1681 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     1741 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     1801 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     1861 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1921 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1981 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2041 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     2101 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2161 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2221 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2281 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2341 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2401 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2461 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2521 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2581 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     2641 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     2701 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     2761 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     2821 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     2881 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     2941 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     3001 ggcagaacgc catcaaaaat aattcgcgtc tggccttcct gtagccagct ttcatcaaca
     3061 ttaaatgtga gcgagtaaca acccgtcgga ttctccgtgg gaacaaacgg cggattgacc
     3121 gtaatgggat aggtcacgtt ggtgtagatg ggcgcatcgt aaccgtgcat ctgccagttt
     3181 gaggggacga cgacagtatc ggcctcagga agatcgcact ccagccagct ttccggcacc
     3241 gcttctggtg ccggaaacca ggcaaagcgc cattcgccat tcaggctgcg caactgttgg
     3301 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     3361 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     3421 gccagtgaat ccgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     3481 aattccacac attatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3541 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3601 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3661 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     3721 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     3781 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
     3841 ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     3901 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     3961 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     4021 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     4081 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     4141 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
     4201 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     4261 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     4321 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     4381 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     4441 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     4501 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     4561 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     4621 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     4681 ggcgctatca tgccataccg cgaaaggttt tgcaccattc gatggtgtcc tggcacgaca
     4741 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagctcactc
     4801 attaggcacc ccaggcttta cactttatgc ttccggctcg tataatgtgt ggaattgtga
     4861 gcggataaca atttcacaca ggaaacagga tcgatccatc gatgagctta ctccccatcc
     4921 ccctgttgac aattaatcat cggctcgtat aatgtgtgga attgtgagcg gataacaatt
     4981 tcacacagga aacaggatca gcttactccc catccccctg ttgacaatta atcatcggct
     5041 cgtataatgt gtggaattgt gagcggataa caatttcaca caggaaaca
//
