LOCUS       sequence-418259-       10096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector for Type 0 promoter 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  JOURNAL   unpublished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            "unpublished"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-P_cRed_BsaI
FEATURES             Location/Qualifiers
     source          1..10096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(374..468)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(489..1148)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1063..1082
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1310..1328
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1332..1375)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     regulatory      7486..7495
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      9371..9418
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      9447..9504
                     /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(9528..9545)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(9699..10096,1..191))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9779..9798)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
       61 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
      121 aaaccaccgc taccagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
      181 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
      241 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
      301 taaattaaaa attaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
      361 gctcgaggct tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca
      421 aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg
      481 atatcaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt
      541 ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc
      601 accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc
      661 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgacacgaaa
      721 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
      781 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
      841 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
      901 accagctcac cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca
      961 agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag
     1021 gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc
     1081 tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt
     1141 ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt
     1201 agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgcccgat caactcgagt
     1261 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     1321 cacgaggcag aatttcagat aaaaaaaatc cttagctttc gctaaggatg atttcgactt
     1381 ggtctcgcta ttgagaccgc agcggtctgc caatccgacg gaggtttatg aattttccac
     1441 cttttccaca agctcaacta gtattaacga tgtggattta gcaaaaaaaa cctgtaaccc
     1501 taaatgtaaa ataacgggta agcctgccaa ccatgttatg gcagattaag cgtctttttg
     1561 aagggcaccg catctttcgc gttgccgtaa atgtatccgt ttagcaataa ggaaattctt
     1621 aatgaataat ccgtcgttac tcaatcatgc ggtcgaaacg atggcagttg gctcgaaaag
     1681 ttttgcgaca gcctcaaagt tatttgatgc aaaaacccgg cgcagcgtac tgatgctcta
     1741 cgcctggtgc cgccattgtg acgatgttat tgacgatcag acgctgggct ttcaggcccg
     1801 gcagcctgcc ttacaaacgc ccgaacaacg tctgatgcaa cttgagatga aaacgcgcca
     1861 ggcctatgca ggatcgcaga tgcacgaacc ggcgtttgcg gcttttcggg aagtggctat
     1921 ggctcatgat atcgccccgg cttacgcgtt tgatcatctg gaaggcttcg ccatggatgt
     1981 acgcgaagcg caatacagcc aactggatga tacgctgcgc tattgctatc acgttgcagg
     2041 cgttgtcggc ttgatgatgg cgcaaatcat gggcgtgcgg gataacgcca cgctggaccg
     2101 cgcctgtgac cttgggctgg catttcagtt gaccaatatt gctcgcgata ttgtggacga
     2161 tgcgcatgcg ggccgctgtt atctgccggc aagctggctg gagcatgaag gtctgaacaa
     2221 agagaattat gcggcacctg aaaaccgtca ggcgctgagc cgtatcgccc gtcgtttggt
     2281 gcaggaagca gaaccttact atttgtctgc cacagccggc ctggcagggt tgcccctgcg
     2341 ttccgcctgg gcaatcgcta cggcgaagca ggtttaccgg aaaataggtg tcaaagttga
     2401 acaggccggt cagcaagcct gggatcagcg gcagtcaacg accacgcccg aaaaattaac
     2461 gctgctgctg gccgcctctg gtcaggccct tacttcccgg atgcgggctc atcctccccg
     2521 ccctgcgcat ctctggcagc gcccgctcta ggcttactat aaggatgttt gcaatgaaac
     2581 caactacggt aattggtgca ggcttcggtg gcctggcact ggcaattcgt ctacaagctg
     2641 cggggatccc cgtcttactg cttgaacaac gtgataaacc cggcggtcgg gcttatgtct
     2701 acgaggatca ggggtttacc tttgatgcag gcccgacggt tatcaccgat cccagtgcca
     2761 ttgaagaact gtttgcactg gcaggaaaac agttaaaaga gtatgtcgaa ctgctgccgg
     2821 ttacgccgtt ttaccgcctg tgttgggagt cagggaaggt ctttaattac gataacgatc
     2881 aaacccggct cgaagcgcag attcagcagt ttaatccccg cgatgtcgaa ggttatcgtc
     2941 agtttctgga ctattcacgc gcggtgttta aagaaggcta tctaaagctc ggtactgtcc
     3001 cttttttatc gttcagagac atgcttcgcg ccgcacctca actggcgaaa ctgcaggcat
     3061 ggagaagcgt ttacagtaag gttgccagtt acatcgaaga tgaacatctg cgccaggcgt
     3121 tttctttcca ctcgctgttg gtgggcggca atcccttcgc cacctcatcc atttatacgt
     3181 tgatacacgc gctggagcgt gagtggggcg tctggtttcc gcgtggcggc accggcgcat
     3241 tagttcaggg gatgataaag ctgtttcagg atctgggtgg cgaagtcgtg ttaaacgcca
     3301 gagtcagcca tatggaaacg acaggaaaca agattgaagc cgtgcattta gaggacggtc
     3361 gcaggttcct gacgcaagcc gtcgcgtcaa atgcagatgt ggttcatacc tatcgcgacc
     3421 tgttaagcca gcaccctgcc gcggttaagc agtccaacaa actgcagact aagcgcatga
     3481 gtaactctct gtttgtgctc tattttggtt tgaatcacca tcatgatcag ctcgcgcatc
     3541 acacggtttg tttcggcccg cgttaccgcg agctgattga cgaaattttt aatcatgatg
     3601 gcctcgcaga ggacttctca ctttatctgc acgcgccctg tgtcacggat tcgtcactgg
     3661 cgcctgaagg ttgcggcagt tactatgtgt tggcgccggt gccgcattta ggcaccgcga
     3721 acctcgactg gacggttgag gggccaaaac tacgcgaccg tatttttgcg taccttgagc
     3781 agcattacat gcctggctta cggagtcagc tggtcacgca ccggatgttt acgccgtttg
     3841 attttcgcga ccagcttaat gcctatcatg gctcagcctt ttctgtggag cccgttctta
     3901 cccagagcgc ctggtttcgg ccgcataacc gcgataaaac cattactaat ctctacctgg
     3961 tcggcgcagg cacgcatccc ggcgcaggca ttcctggcgt catcggctcg gcaaaagcga
     4021 cagcaggttt gatgctggag gatctgatat gagcttttac ccaggagaca gagaatgcaa
     4081 ccgcattatg atctgattct cgtgggggct ggactcgcga atggccttat cgccctgcga
     4141 cttcagcagc agcaacctga tatgcgtatt ttgcttatcg acgccgcacc ccaggcgggc
     4201 gggaatcata cgtggtcatt tcaccacgat gatttgactg agagccaaca tcgttggata
     4261 gctccgctgg tggttcatca ctggcccgac tatcaggtac gctttcccac acgccgtcgt
     4321 aagctgaaca gcggctactt ttgtattact tctcagcgtt tcgctgaggt tttacagcga
     4381 cagtttggcc cgcacttgtg gatggatacc gcggtcgcag aggttaatgc ggaatctgtt
     4441 cggttgaaaa agggtcaggt tatcggtgcc cgcgcggtga ttgacgggcg gggttatgcg
     4501 gcaaattcag cactgagcgt gggcttccag gcgtttattg gccaggaatg gcgattgagt
     4561 cacccgcatg gtttatcatc tccgattatc atggatgcca cggtcgatca gcaaaatggt
     4621 tatcgcttcg tgtacagcct gccgctctcg ccgaccagat tgttaattga agatacgcac
     4681 tatattgata atgcgacatt agatcctgaa tgcgcgcggc aaaatatttg cgactatgcc
     4741 gcgcaacagg gttggcagct tcagacactg ctgcgagaag aacagggcgc cttacccatt
     4801 actctgtcgg gcaatgccga cgcattctgg cagcagcgcc ccctggcctg tagtggatta
     4861 cgtgccggtc tgttccatcc taccaccggc tattcactgc cgctggcggt tgccgtggcc
     4921 gaccgcctga gtgcacttga tgtctttacg tcggcctcaa ttcaccatgc cattacgcat
     4981 tttgcccgcg agcgctggca gcagcagggc tttttccgca tgctgaatcg catgctgttt
     5041 ttagccggac ccgccgattc acgctggcgg gttatgcagc gtttttatgg tttacctgaa
     5101 gatttaattg cccgttttta tgcgggaaaa ctcacgctga ccgatcggct acgtattctg
     5161 agcggcaagc cgcctgttcc ggtattagca gcattgcaag ccattatgac gactcatcgt
     5221 taagcttcag aggaggaatt aagtaatgag ttttgatatt gccaaatacc cgaccctggc
     5281 actggtcgac tccacccagg agttacgact gttgccgaaa gagagtttac cgaaactctg
     5341 cgacgaactg cgccgctatt tactcgacag cgtgagccgt tccagcgggc acttcgcctc
     5401 cgggctgggc acggtcgaac tgaccgtggc gctgcactat gtctacaaca ccccgtttga
     5461 ccaattgatt tgggatgtgg ggcatcaggc ttatccgcat aaaattttga ccggacgccg
     5521 cgacaaaatc ggcaccatcc gtcagaaagg cggtctgcac ccgttcccgt ggcgcggcga
     5581 aagcgaatat gacgtattaa gcgtcgggca ttcatcaacc tccattagtg ccggaattgg
     5641 tattgcggtt gctgccgaaa aagaaggcaa aaatcgccgc accgtctgtg tcattggcga
     5701 tggcgcgatt accgcaggca tggcgtttga agcgatgaat cacgcgggcg atatccgtcc
     5761 tgatatgctg gtgattctca acgacaatga aatgtcgatt tccgaaaatg tcggcgcgct
     5821 caacaaccat ctggcacagc tgctttccgg taagctttac tcttcactgc gcgaaggcgg
     5881 gaaaaaagtt ttctctggcg tgccgccaat taaagagctg ctcaaacgca ccgaagaaca
     5941 tattaaaggc atggtagtgc ctggcacgtt gtttgaggag ctgggcttta actacatcgg
     6001 cccggtggac ggtcatgatg tgctggggct tatcaccacg ctaaagaaca tgcgcgacct
     6061 gaaaggcccg cagttcctgc atatcatgac caaaaaaggt cgtggttatg aaccggcaga
     6121 aaaagacccg atcactttcc atgccgtgct aaatttgatc cctccagcgg ttgtttgccg
     6181 aaaagtagcg gcggtttgcc gagctattca aaaatctttg gcgactggtt gtgcgaaacg
     6241 gcagcgaaag acaacaagct gatggcgatt actccggcga tgcgtgaagg ttccggcatg
     6301 gtcgagtttt cacgtaaatt cccggatcgc tacttcgacg tggcaattgc cgagcaacat
     6361 gcggtgacct ttgctgcggg tctggcgatt ggtgggtaca aacccattgt cgcgatttac
     6421 tccactttcc tgcaacgcgc ctatgatcag gtgctgcatg acgtggcgat tcaaaagctt
     6481 ccggtcctgt tcgccatcga ccgcgcgggc attgttggtg ctgacggtca aacccatcag
     6541 ggtgcttttg atctctctta cctgcgctgc ataccggaaa tggtcattat gaccccgagc
     6601 gatgaaaacg aatgtcgcca gatgctctat accggctatc actataacga tggcccgtca
     6661 gcggtgcgct acccgcgtgg caacgcggtc ggcgtggaac tgacgccgct ggaaaaacta
     6721 ccaattggca aaggcattgt gaagcgtcgt ggcgagaaac tggcgatcct taactttggt
     6781 acgctgatgc cagaagcggc gaaagtcgcc aaatcgctga acgccacgct ggtcgatatg
     6841 cgttttgtga aaccgcttga tgaagcgtta attctggaaa tggccgccag ccatgaagcg
     6901 ctggtcaccg tagaagaaaa cgccattatg ggcggcgcag gcagcggcgt gaacgaagtg
     6961 ctgatggccc atcgtaaacc agtacccgtg ctgaacattg gcctgccgga cttctttatt
     7021 ccgcaaggaa ctcaggaaga aatgcgcgcc gaactcggcc tcgatgccgc tggtatggaa
     7081 gccaaaatca aggcctggct ggcataagct ttgtgtgagg agatgtcgaa tgagcgcaca
     7141 tgccctgccc aaggcagacc tgaccgccac cagcctgatc gtatcgggcg gcatcatcgc
     7201 cgcttggctg gccctgcatg tgcatgcgct gtggtttctg gacgcagcgg cgcatcccat
     7261 cctggcgatc gcaaatttcc tggggctgac ctggctgtcg gtcggattgt tcatcatcgc
     7321 gcatgacgcg atgcacgggt cggtggtgcc ggggcgtccg cgcgccaatg cggcgatggg
     7381 ccagcttgtc ctgtggctgt atgccggatt ttcgtggcgc aagatgatcg tcaagcacat
     7441 ggcccatcac cgccatgccg gaaccgacga cgaccccgat ttcgaccatg gcggcccggt
     7501 ccgctggtac gcccgcttca tcggcaccta tttcggctgg cgcgaggggc tgcttctgcc
     7561 cgtcatcgtg acggtctatg cgctgatcct tggggatcgc tggatgtacg tggtattctg
     7621 gccgctgccg tcgatcctgg cgtcgatcca gctgttcgtg ttcggcacct ggctgccgca
     7681 ccgccccggc cacgacgcgt tcccggaccg ccacaatgcg cggtcgtcgc ggatcagcga
     7741 ccccgtgtcg ctgctgacct gttttcactt tggcggttat catcacgaac accacctgca
     7801 cccgacggtg ccgtggtggc gcctgcccag cacccgcacc aagggcgaca ccgcataagc
     7861 ttgagctgag gagattttga atgcaaacgg aacacgtcat tttattgaat gcacagggag
     7921 ttcccacggg tacgctggaa aagtatgccg cacacacggc agacacccgc ttacatctcg
     7981 cgttctccag ttggctgttt aatgccaaag gacaattatt agttacccgc cgcgcactga
     8041 gcaaaaaagc atggcctggc gtgtggacta actcggtttg tgggcaccca caactgggag
     8101 aaagcaacga ggacgcagtg atccgccgtt gccgttatga gcttggcgtg gaaattacgc
     8161 ctcctgaatc tatctatcct gactttcgct accgcgccac cgatccgagt ggcattgtgg
     8221 aaaatgaagt gtgtccggta tttgccgcac gcaccactag tgcgttacag atcaatgatg
     8281 atgaagtgat ggattatcaa tggtgtgatt tagcagatgt attacacggt attgatgcaa
     8341 cgccgtgggc gttcagtccg tggatggtga tgcaggcgac aaatcgcgaa gccagaaaac
     8401 gattatctgc atttacccag cttaaataag ctttgcctca ggatttttga aatgacggtc
     8461 tgcgcaaaaa aacacgttca tctcactcgc gatgctgcgg agcagttact ggctgatatt
     8521 gatcgacgcc ttgatcagtt attgcccgtg gagggagaac gggatgttgt gggtgccgcg
     8581 atgcgtgaag gtgcgctggc accgggaaaa cgtattcgcc ccatgttgct gttgctgacc
     8641 gcccgcgatc tgggttgcgc tttcagccat gacggattac tggatttggc ctgtgcggtg
     8701 gaaatggtcc acgcggcttc gctgatcctt gacgatatgc cctgcatgga cgatgcgaag
     8761 ctgcggcgcg gacgccctac cattcattct cattacggag agcatgtggc aatactggcg
     8821 gcggttgcct tgctgagtaa agcctttggc gtaattgccg atgcagatgg cctcacgccg
     8881 ctggcaaaaa atcgggcggt ttctgaactg tcaaacgcca tcggcatgca aggattggtt
     8941 cagggtcagt tcaaggatct gtctgaaggg gataagccgc gcagcgctga agctattttg
     9001 atgacgaatc actttaaaac cagcacgctg ttttgtgcct cgatgcagat ggcctcgatt
     9061 gttgcgaatg cctccagcga agcgcgtgat tgcctgcatc gtttttcact tgatcttggt
     9121 caggcatttc aactgctgga cgatttgacc gatggcatga ccgacaccgg taaggatagc
     9181 aatcaggacg ccggtaaatc gacgctggtc aatctgttag gcccgagggc ggttgaagaa
     9241 cgtctgagac aacatcttca gcttgccagt gagcatctct ctgcggcctg ccaacacggg
     9301 cacgccactc aacattttat tcaggcctgg tttgacaaaa aactcgctgc cgtcagttaa
     9361 gcttgcaaga ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttggt
     9421 gacggtctca gtaccgagac caagtctccg gcaaaaaagg gcaaggtgtc accaccctgc
     9481 cctttttctt taaaaccgaa aagattactt cgcgttatgc aggcttcctc gctcactgac
     9541 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     9601 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     9661 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccacaggct ccgcccccct
     9721 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     9781 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     9841 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     9901 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     9961 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
    10021 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
    10081 tatgtaggcg gtgcta
//
