LOCUS       sequence-418267-       10096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector for Type 0 terminator parts.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10096)
  AUTHORS   Shapiro M
  TITLE     EcoFlex MoClo Extension, Volume I
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 10096)
  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            "unpublished"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: pMetBP-T_cRed_BsaI
FEATURES             Location/Qualifiers
     source          1..10096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(163..257)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(278..937)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     852..871
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1099..1117
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1121..1164)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     regulatory      7275..7284
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      9160..9207
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      9236..9293
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(9317..9334)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9488..10076)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9568..9587)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
       61 atcaaaaagg atcttcacct agatcctttt aaattaaaaa ttaagtttta aatcaatcta
      121 aagtatatat gagtaaactt ggtctgacag ctcgaggctt ggattctcac caataaaaaa
      181 cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg
      241 atctatcaac aggagtccaa gcgagctcga tatcaaatta cgccccgccc tgccactcat
      301 cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat
      361 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
      421 tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga
      481 aactcaccca gggattggct gacacgaaaa acatattctc aataaaccct ttagggaaat
      541 aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga
      601 aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg
      661 tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgaa
      721 attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt
      781 gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat
      841 aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata
      901 tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa
      961 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
     1021 aacctcttac gtgcccgatc aactcgagtg ccacctgacg tctaagaaac cattattatc
     1081 atgacattaa cctataaaaa taggcgtatc acgaggcaga atttcagata aaaaaaatcc
     1141 ttagctttcg ctaaggatga tttcgacttg gtctcgtcga tgagaccgca gcggtctgcc
     1201 aatccgacgg aggtttatga attttccacc ttttccacaa gctcaactag tattaacgat
     1261 gtggatttag caaaaaaaac ctgtaaccct aaatgtaaaa taacgggtaa gcctgccaac
     1321 catgttatgg cagattaagc gtctttttga agggcaccgc atctttcgcg ttgccgtaaa
     1381 tgtatccgtt tagcaataag gaaattctta atgaataatc cgtcgttact caatcatgcg
     1441 gtcgaaacga tggcagttgg ctcgaaaagt tttgcgacag cctcaaagtt atttgatgca
     1501 aaaacccggc gcagcgtact gatgctctac gcctggtgcc gccattgtga cgatgttatt
     1561 gacgatcaga cgctgggctt tcaggcccgg cagcctgcct tacaaacgcc cgaacaacgt
     1621 ctgatgcaac ttgagatgaa aacgcgccag gcctatgcag gatcgcagat gcacgaaccg
     1681 gcgtttgcgg cttttcggga agtggctatg gctcatgata tcgccccggc ttacgcgttt
     1741 gatcatctgg aaggcttcgc catggatgta cgcgaagcgc aatacagcca actggatgat
     1801 acgctgcgct attgctatca cgttgcaggc gttgtcggct tgatgatggc gcaaatcatg
     1861 ggcgtgcggg ataacgccac gctggaccgc gcctgtgacc ttgggctggc atttcagttg
     1921 accaatattg ctcgcgatat tgtggacgat gcgcatgcgg gccgctgtta tctgccggca
     1981 agctggctgg agcatgaagg tctgaacaaa gagaattatg cggcacctga aaaccgtcag
     2041 gcgctgagcc gtatcgcccg tcgtttggtg caggaagcag aaccttacta tttgtctgcc
     2101 acagccggcc tggcagggtt gcccctgcgt tccgcctggg caatcgctac ggcgaagcag
     2161 gtttaccgga aaataggtgt caaagttgaa caggccggtc agcaagcctg ggatcagcgg
     2221 cagtcaacga ccacgcccga aaaattaacg ctgctgctgg ccgcctctgg tcaggccctt
     2281 acttcccgga tgcgggctca tcctccccgc cctgcgcatc tctggcagcg cccgctctag
     2341 gcttactata aggatgtttg caatgaaacc aactacggta attggtgcag gcttcggtgg
     2401 cctggcactg gcaattcgtc tacaagctgc ggggatcccc gtcttactgc ttgaacaacg
     2461 tgataaaccc ggcggtcggg cttatgtcta cgaggatcag gggtttacct ttgatgcagg
     2521 cccgacggtt atcaccgatc ccagtgccat tgaagaactg tttgcactgg caggaaaaca
     2581 gttaaaagag tatgtcgaac tgctgccggt tacgccgttt taccgcctgt gttgggagtc
     2641 agggaaggtc tttaattacg ataacgatca aacccggctc gaagcgcaga ttcagcagtt
     2701 taatccccgc gatgtcgaag gttatcgtca gtttctggac tattcacgcg cggtgtttaa
     2761 agaaggctat ctaaagctcg gtactgtccc ttttttatcg ttcagagaca tgcttcgcgc
     2821 cgcacctcaa ctggcgaaac tgcaggcatg gagaagcgtt tacagtaagg ttgccagtta
     2881 catcgaagat gaacatctgc gccaggcgtt ttctttccac tcgctgttgg tgggcggcaa
     2941 tcccttcgcc acctcatcca tttatacgtt gatacacgcg ctggagcgtg agtggggcgt
     3001 ctggtttccg cgtggcggca ccggcgcatt agttcagggg atgataaagc tgtttcagga
     3061 tctgggtggc gaagtcgtgt taaacgccag agtcagccat atggaaacga caggaaacaa
     3121 gattgaagcc gtgcatttag aggacggtcg caggttcctg acgcaagccg tcgcgtcaaa
     3181 tgcagatgtg gttcatacct atcgcgacct gttaagccag caccctgccg cggttaagca
     3241 gtccaacaaa ctgcagacta agcgcatgag taactctctg tttgtgctct attttggttt
     3301 gaatcaccat catgatcagc tcgcgcatca cacggtttgt ttcggcccgc gttaccgcga
     3361 gctgattgac gaaattttta atcatgatgg cctcgcagag gacttctcac tttatctgca
     3421 cgcgccctgt gtcacggatt cgtcactggc gcctgaaggt tgcggcagtt actatgtgtt
     3481 ggcgccggtg ccgcatttag gcaccgcgaa cctcgactgg acggttgagg ggccaaaact
     3541 acgcgaccgt atttttgcgt accttgagca gcattacatg cctggcttac ggagtcagct
     3601 ggtcacgcac cggatgttta cgccgtttga ttttcgcgac cagcttaatg cctatcatgg
     3661 ctcagccttt tctgtggagc ccgttcttac ccagagcgcc tggtttcggc cgcataaccg
     3721 cgataaaacc attactaatc tctacctggt cggcgcaggc acgcatcccg gcgcaggcat
     3781 tcctggcgtc atcggctcgg caaaagcgac agcaggtttg atgctggagg atctgatatg
     3841 agcttttacc caggagacag agaatgcaac cgcattatga tctgattctc gtgggggctg
     3901 gactcgcgaa tggccttatc gccctgcgac ttcagcagca gcaacctgat atgcgtattt
     3961 tgcttatcga cgccgcaccc caggcgggcg ggaatcatac gtggtcattt caccacgatg
     4021 atttgactga gagccaacat cgttggatag ctccgctggt ggttcatcac tggcccgact
     4081 atcaggtacg ctttcccaca cgccgtcgta agctgaacag cggctacttt tgtattactt
     4141 ctcagcgttt cgctgaggtt ttacagcgac agtttggccc gcacttgtgg atggataccg
     4201 cggtcgcaga ggttaatgcg gaatctgttc ggttgaaaaa gggtcaggtt atcggtgccc
     4261 gcgcggtgat tgacgggcgg ggttatgcgg caaattcagc actgagcgtg ggcttccagg
     4321 cgtttattgg ccaggaatgg cgattgagtc acccgcatgg tttatcatct ccgattatca
     4381 tggatgccac ggtcgatcag caaaatggtt atcgcttcgt gtacagcctg ccgctctcgc
     4441 cgaccagatt gttaattgaa gatacgcact atattgataa tgcgacatta gatcctgaat
     4501 gcgcgcggca aaatatttgc gactatgccg cgcaacaggg ttggcagctt cagacactgc
     4561 tgcgagaaga acagggcgcc ttacccatta ctctgtcggg caatgccgac gcattctggc
     4621 agcagcgccc cctggcctgt agtggattac gtgccggtct gttccatcct accaccggct
     4681 attcactgcc gctggcggtt gccgtggccg accgcctgag tgcacttgat gtctttacgt
     4741 cggcctcaat tcaccatgcc attacgcatt ttgcccgcga gcgctggcag cagcagggct
     4801 ttttccgcat gctgaatcgc atgctgtttt tagccggacc cgccgattca cgctggcggg
     4861 ttatgcagcg tttttatggt ttacctgaag atttaattgc ccgtttttat gcgggaaaac
     4921 tcacgctgac cgatcggcta cgtattctga gcggcaagcc gcctgttccg gtattagcag
     4981 cattgcaagc cattatgacg actcatcgtt aagcttcaga ggaggaatta agtaatgagt
     5041 tttgatattg ccaaataccc gaccctggca ctggtcgact ccacccagga gttacgactg
     5101 ttgccgaaag agagtttacc gaaactctgc gacgaactgc gccgctattt actcgacagc
     5161 gtgagccgtt ccagcgggca cttcgcctcc gggctgggca cggtcgaact gaccgtggcg
     5221 ctgcactatg tctacaacac cccgtttgac caattgattt gggatgtggg gcatcaggct
     5281 tatccgcata aaattttgac cggacgccgc gacaaaatcg gcaccatccg tcagaaaggc
     5341 ggtctgcacc cgttcccgtg gcgcggcgaa agcgaatatg acgtattaag cgtcgggcat
     5401 tcatcaacct ccattagtgc cggaattggt attgcggttg ctgccgaaaa agaaggcaaa
     5461 aatcgccgca ccgtctgtgt cattggcgat ggcgcgatta ccgcaggcat ggcgtttgaa
     5521 gcgatgaatc acgcgggcga tatccgtcct gatatgctgg tgattctcaa cgacaatgaa
     5581 atgtcgattt ccgaaaatgt cggcgcgctc aacaaccatc tggcacagct gctttccggt
     5641 aagctttact cttcactgcg cgaaggcggg aaaaaagttt tctctggcgt gccgccaatt
     5701 aaagagctgc tcaaacgcac cgaagaacat attaaaggca tggtagtgcc tggcacgttg
     5761 tttgaggagc tgggctttaa ctacatcggc ccggtggacg gtcatgatgt gctggggctt
     5821 atcaccacgc taaagaacat gcgcgacctg aaaggcccgc agttcctgca tatcatgacc
     5881 aaaaaaggtc gtggttatga accggcagaa aaagacccga tcactttcca tgccgtgcta
     5941 aatttgatcc ctccagcggt tgtttgccga aaagtagcgg cggtttgccg agctattcaa
     6001 aaatctttgg cgactggttg tgcgaaacgg cagcgaaaga caacaagctg atggcgatta
     6061 ctccggcgat gcgtgaaggt tccggcatgg tcgagttttc acgtaaattc ccggatcgct
     6121 acttcgacgt ggcaattgcc gagcaacatg cggtgacctt tgctgcgggt ctggcgattg
     6181 gtgggtacaa acccattgtc gcgatttact ccactttcct gcaacgcgcc tatgatcagg
     6241 tgctgcatga cgtggcgatt caaaagcttc cggtcctgtt cgccatcgac cgcgcgggca
     6301 ttgttggtgc tgacggtcaa acccatcagg gtgcttttga tctctcttac ctgcgctgca
     6361 taccggaaat ggtcattatg accccgagcg atgaaaacga atgtcgccag atgctctata
     6421 ccggctatca ctataacgat ggcccgtcag cggtgcgcta cccgcgtggc aacgcggtcg
     6481 gcgtggaact gacgccgctg gaaaaactac caattggcaa aggcattgtg aagcgtcgtg
     6541 gcgagaaact ggcgatcctt aactttggta cgctgatgcc agaagcggcg aaagtcgcca
     6601 aatcgctgaa cgccacgctg gtcgatatgc gttttgtgaa accgcttgat gaagcgttaa
     6661 ttctggaaat ggccgccagc catgaagcgc tggtcaccgt agaagaaaac gccattatgg
     6721 gcggcgcagg cagcggcgtg aacgaagtgc tgatggccca tcgtaaacca gtacccgtgc
     6781 tgaacattgg cctgccggac ttctttattc cgcaaggaac tcaggaagaa atgcgcgccg
     6841 aactcggcct cgatgccgct ggtatggaag ccaaaatcaa ggcctggctg gcataagctt
     6901 tgtgtgagga gatgtcgaat gagcgcacat gccctgccca aggcagacct gaccgccacc
     6961 agcctgatcg tatcgggcgg catcatcgcc gcttggctgg ccctgcatgt gcatgcgctg
     7021 tggtttctgg acgcagcggc gcatcccatc ctggcgatcg caaatttcct ggggctgacc
     7081 tggctgtcgg tcggattgtt catcatcgcg catgacgcga tgcacgggtc ggtggtgccg
     7141 gggcgtccgc gcgccaatgc ggcgatgggc cagcttgtcc tgtggctgta tgccggattt
     7201 tcgtggcgca agatgatcgt caagcacatg gcccatcacc gccatgccgg aaccgacgac
     7261 gaccccgatt tcgaccatgg cggcccggtc cgctggtacg cccgcttcat cggcacctat
     7321 ttcggctggc gcgaggggct gcttctgccc gtcatcgtga cggtctatgc gctgatcctt
     7381 ggggatcgct ggatgtacgt ggtattctgg ccgctgccgt cgatcctggc gtcgatccag
     7441 ctgttcgtgt tcggcacctg gctgccgcac cgccccggcc acgacgcgtt cccggaccgc
     7501 cacaatgcgc ggtcgtcgcg gatcagcgac cccgtgtcgc tgctgacctg ttttcacttt
     7561 ggcggttatc atcacgaaca ccacctgcac ccgacggtgc cgtggtggcg cctgcccagc
     7621 acccgcacca agggcgacac cgcataagct tgagctgagg agattttgaa tgcaaacgga
     7681 acacgtcatt ttattgaatg cacagggagt tcccacgggt acgctggaaa agtatgccgc
     7741 acacacggca gacacccgct tacatctcgc gttctccagt tggctgttta atgccaaagg
     7801 acaattatta gttacccgcc gcgcactgag caaaaaagca tggcctggcg tgtggactaa
     7861 ctcggtttgt gggcacccac aactgggaga aagcaacgag gacgcagtga tccgccgttg
     7921 ccgttatgag cttggcgtgg aaattacgcc tcctgaatct atctatcctg actttcgcta
     7981 ccgcgccacc gatccgagtg gcattgtgga aaatgaagtg tgtccggtat ttgccgcacg
     8041 caccactagt gcgttacaga tcaatgatga tgaagtgatg gattatcaat ggtgtgattt
     8101 agcagatgta ttacacggta ttgatgcaac gccgtgggcg ttcagtccgt ggatggtgat
     8161 gcaggcgaca aatcgcgaag ccagaaaacg attatctgca tttacccagc ttaaataagc
     8221 tttgcctcag gatttttgaa atgacggtct gcgcaaaaaa acacgttcat ctcactcgcg
     8281 atgctgcgga gcagttactg gctgatattg atcgacgcct tgatcagtta ttgcccgtgg
     8341 agggagaacg ggatgttgtg ggtgccgcga tgcgtgaagg tgcgctggca ccgggaaaac
     8401 gtattcgccc catgttgctg ttgctgaccg cccgcgatct gggttgcgct ttcagccatg
     8461 acggattact ggatttggcc tgtgcggtgg aaatggtcca cgcggcttcg ctgatccttg
     8521 acgatatgcc ctgcatggac gatgcgaagc tgcggcgcgg acgccctacc attcattctc
     8581 attacggaga gcatgtggca atactggcgg cggttgcctt gctgagtaaa gcctttggcg
     8641 taattgccga tgcagatggc ctcacgccgc tggcaaaaaa tcgggcggtt tctgaactgt
     8701 caaacgccat cggcatgcaa ggattggttc agggtcagtt caaggatctg tctgaagggg
     8761 ataagccgcg cagcgctgaa gctattttga tgacgaatca ctttaaaacc agcacgctgt
     8821 tttgtgcctc gatgcagatg gcctcgattg ttgcgaatgc ctccagcgaa gcgcgtgatt
     8881 gcctgcatcg tttttcactt gatcttggtc aggcatttca actgctggac gatttgaccg
     8941 atggcatgac cgacaccggt aaggatagca atcaggacgc cggtaaatcg acgctggtca
     9001 atctgttagg cccgagggcg gttgaagaac gtctgagaca acatcttcag cttgccagtg
     9061 agcatctctc tgcggcctgc caacacgggc acgccactca acattttatt caggcctggt
     9121 ttgacaaaaa actcgctgcc gtcagttaag cttgcaagac tagcataacc ccttggggcc
     9181 tctaaacggg tcttgagggg ttttttggtg acggtctcat gttcgagacc aagtctccgg
     9241 caaaaaaggg caaggtgtca ccaccctgcc ctttttcttt aaaaccgaaa agattacttc
     9301 gcgttatgca ggcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     9361 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     9421 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     9481 tggcgttttt ccacaggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     9541 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     9601 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     9661 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     9721 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     9781 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     9841 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     9901 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     9961 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctacca
    10021 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
    10081 atcctttgat cttttc
//
