LOCUS       sequence-174143-        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 1
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 WT*
FEATURES             Location/Qualifiers
     source          1..5089
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(22..27,28..45,46..52)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    60..76
                     /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(131..136,137..152,153..159)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    167..183
                     /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(226..231,232..247,248..254)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    262..278
                     /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(1022..1040)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1108..1212
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1213..1281,1282..2073)
                     /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(1431..1450)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2244..2832
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2733..2752
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2986..3003
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3018..3158
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3084..3103)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3244..3266
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3279..3740)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVLPRAFR"
     primer_bind     3689..3711
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3703..3720
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3704..3720
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3736..3752)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3736..3752)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3760..3776
                     /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(3784..3790,3791..3808,3809..3814))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             complement(3863..4945)
                     /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     4907..4926
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4946..5023)
                     /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."
ORIGIN
        1 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggaat
       61 tgtgagcgga taacaatttc acacaggaaa caggatcgat ccatcgatga gcttactccc
      121 catccccctg ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa
      181 caatttcaca caggaaacag gatcagctta ctccccatcc ccctgttgac aattaatcat
      241 cggctcgtat aatgtgtgga attgtgagcg gataacaatt tcacacagga aacaggatcc
      301 atcgatgctt aggaggtatt atgaatatat ttgaaatgtt acgtatagat gaaggtctta
      361 gacttaaaat ctataaagac acagaaggct attacactat tggcatcggt catttgctta
      421 caaaaagtcc atcacttaat gctgctaaat ctgaattaga taaagctatt gggcgtaata
      481 ctaatggtgt aattacaaaa gatgaggctg aaaaactctt taatcaggat gttgatgctg
      541 ctgttcgcgg aattctgaga aatgctaaat taaaaccggt ttatgattct cttgatgcgg
      601 ttcgtcgcgc tgcattgatt aatatggttt tccaaatggg agaaaccggt gtggcaggat
      661 ttactaactc tttacgtatg cttcaacaaa aacgctggga tgaagcagca gttaacttag
      721 ctaaaagtag atggtataat caaacaccta atcgcgcaaa acgagtcatt acaacgttta
      781 gaactggcac ttgggacgcg tataaaaatc tataaagctg tttactttct cttggaattg
      841 tgatagtata ttcacaatta cttgaataga caattactaa ttaaaatatt taaaggaaac
      901 atatgaaaac atatcaagaa tttattgccg aagcttatcg atgataagct gtcaaacatg
      961 agcagatctg agcccgccta atgagcgggc ttttttttca gatctgcttg aagacgaaag
     1021 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     1081 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     1141 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     1201 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     1261 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     1321 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     1381 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     1441 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     1501 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     1561 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     1621 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     1681 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     1741 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     1801 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     1861 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     1921 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     1981 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     2041 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     2101 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     2161 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     2221 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2281 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2341 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     2401 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2461 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2521 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2581 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2641 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2701 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2761 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2821 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2881 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     2941 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3001 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3061 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     3121 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     3181 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3241 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcaga
     3301 acgccatcaa aaataattcg cgtctggcct tcctgtagcc agctttcatc aacattaaat
     3361 gtgagcgagt aacaacccgt cggattctcc gtgggaacaa acggcggatt gaccgtaatg
     3421 ggataggtca cgttggtgta gatgggcgca tcgtaaccgt gcatctgcca gtttgagggg
     3481 acgacgacag tatcggcctc aggaagatcg cactccagcc agctttccgg caccgcttct
     3541 ggtgccggaa accaggcaaa gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     3601 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     3661 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     3721 gaatccgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3781 acacattata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3841 acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3901 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     3961 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     4021 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     4081 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     4141 agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     4201 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     4261 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     4321 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     4381 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     4441 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     4501 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     4561 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
     4621 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
     4681 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
     4741 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
     4801 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
     4861 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
     4921 catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct
     4981 atcatgccat accgcgaaag gttttgcacc attcgatggt gtcctggcac gacaggtttc
     5041 ccgactggaa agcgggcagt gagcgcaacg caattaatgt aagttagct
//
