LOCUS       sequence-418266-       10096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector for Type 0 combined RBS-CDS 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  JOURNAL   unpublished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            "unpublished"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-RC_cRed_BsaI
FEATURES             Location/Qualifiers
     source          1..10096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(432..526)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(547..1206)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1121..1140
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1368..1386
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1390..1433)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     regulatory      7544..7553
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      9429..9476
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      9505..9562
                     /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(9586..9603)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(9757..10096,1..249))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9837..9856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
       61 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
      121 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      181 accaccgcta ccagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
      241 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
      301 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
      361 aattaaaaat taagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagc
      421 tcgaggcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa
      481 tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctcgat
      541 atcaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct
      601 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
      661 cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat
      721 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg acacgaaaaa
      781 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
      841 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
      901 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
      961 cagctcaccg tctttcattg ccatacgaaa ttccggatga gcattcatca ggcgggcaag
     1021 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     1081 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
     1141 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     1201 ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     1261 tgatcttatt tcattatggt gaaagttgga acctcttacg tgcccgatca actcgagtgc
     1321 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     1381 cgaggcagaa tttcagataa aaaaaatcct tagctttcgc taaggatgat ttcgacttgg
     1441 tctcagtact gagaccgcag cggtctgcca atccgacgga ggtttatgaa ttttccacct
     1501 tttccacaag ctcaactagt attaacgatg tggatttagc aaaaaaaacc tgtaacccta
     1561 aatgtaaaat aacgggtaag cctgccaacc atgttatggc agattaagcg tctttttgaa
     1621 gggcaccgca tctttcgcgt tgccgtaaat gtatccgttt agcaataagg aaattcttaa
     1681 tgaataatcc gtcgttactc aatcatgcgg tcgaaacgat ggcagttggc tcgaaaagtt
     1741 ttgcgacagc ctcaaagtta tttgatgcaa aaacccggcg cagcgtactg atgctctacg
     1801 cctggtgccg ccattgtgac gatgttattg acgatcagac gctgggcttt caggcccggc
     1861 agcctgcctt acaaacgccc gaacaacgtc tgatgcaact tgagatgaaa acgcgccagg
     1921 cctatgcagg atcgcagatg cacgaaccgg cgtttgcggc ttttcgggaa gtggctatgg
     1981 ctcatgatat cgccccggct tacgcgtttg atcatctgga aggcttcgcc atggatgtac
     2041 gcgaagcgca atacagccaa ctggatgata cgctgcgcta ttgctatcac gttgcaggcg
     2101 ttgtcggctt gatgatggcg caaatcatgg gcgtgcggga taacgccacg ctggaccgcg
     2161 cctgtgacct tgggctggca tttcagttga ccaatattgc tcgcgatatt gtggacgatg
     2221 cgcatgcggg ccgctgttat ctgccggcaa gctggctgga gcatgaaggt ctgaacaaag
     2281 agaattatgc ggcacctgaa aaccgtcagg cgctgagccg tatcgcccgt cgtttggtgc
     2341 aggaagcaga accttactat ttgtctgcca cagccggcct ggcagggttg cccctgcgtt
     2401 ccgcctgggc aatcgctacg gcgaagcagg tttaccggaa aataggtgtc aaagttgaac
     2461 aggccggtca gcaagcctgg gatcagcggc agtcaacgac cacgcccgaa aaattaacgc
     2521 tgctgctggc cgcctctggt caggccctta cttcccggat gcgggctcat cctccccgcc
     2581 ctgcgcatct ctggcagcgc ccgctctagg cttactataa ggatgtttgc aatgaaacca
     2641 actacggtaa ttggtgcagg cttcggtggc ctggcactgg caattcgtct acaagctgcg
     2701 gggatccccg tcttactgct tgaacaacgt gataaacccg gcggtcgggc ttatgtctac
     2761 gaggatcagg ggtttacctt tgatgcaggc ccgacggtta tcaccgatcc cagtgccatt
     2821 gaagaactgt ttgcactggc aggaaaacag ttaaaagagt atgtcgaact gctgccggtt
     2881 acgccgtttt accgcctgtg ttgggagtca gggaaggtct ttaattacga taacgatcaa
     2941 acccggctcg aagcgcagat tcagcagttt aatccccgcg atgtcgaagg ttatcgtcag
     3001 tttctggact attcacgcgc ggtgtttaaa gaaggctatc taaagctcgg tactgtccct
     3061 tttttatcgt tcagagacat gcttcgcgcc gcacctcaac tggcgaaact gcaggcatgg
     3121 agaagcgttt acagtaaggt tgccagttac atcgaagatg aacatctgcg ccaggcgttt
     3181 tctttccact cgctgttggt gggcggcaat cccttcgcca cctcatccat ttatacgttg
     3241 atacacgcgc tggagcgtga gtggggcgtc tggtttccgc gtggcggcac cggcgcatta
     3301 gttcagggga tgataaagct gtttcaggat ctgggtggcg aagtcgtgtt aaacgccaga
     3361 gtcagccata tggaaacgac aggaaacaag attgaagccg tgcatttaga ggacggtcgc
     3421 aggttcctga cgcaagccgt cgcgtcaaat gcagatgtgg ttcataccta tcgcgacctg
     3481 ttaagccagc accctgccgc ggttaagcag tccaacaaac tgcagactaa gcgcatgagt
     3541 aactctctgt ttgtgctcta ttttggtttg aatcaccatc atgatcagct cgcgcatcac
     3601 acggtttgtt tcggcccgcg ttaccgcgag ctgattgacg aaatttttaa tcatgatggc
     3661 ctcgcagagg acttctcact ttatctgcac gcgccctgtg tcacggattc gtcactggcg
     3721 cctgaaggtt gcggcagtta ctatgtgttg gcgccggtgc cgcatttagg caccgcgaac
     3781 ctcgactgga cggttgaggg gccaaaacta cgcgaccgta tttttgcgta ccttgagcag
     3841 cattacatgc ctggcttacg gagtcagctg gtcacgcacc ggatgtttac gccgtttgat
     3901 tttcgcgacc agcttaatgc ctatcatggc tcagcctttt ctgtggagcc cgttcttacc
     3961 cagagcgcct ggtttcggcc gcataaccgc gataaaacca ttactaatct ctacctggtc
     4021 ggcgcaggca cgcatcccgg cgcaggcatt cctggcgtca tcggctcggc aaaagcgaca
     4081 gcaggtttga tgctggagga tctgatatga gcttttaccc aggagacaga gaatgcaacc
     4141 gcattatgat ctgattctcg tgggggctgg actcgcgaat ggccttatcg ccctgcgact
     4201 tcagcagcag caacctgata tgcgtatttt gcttatcgac gccgcacccc aggcgggcgg
     4261 gaatcatacg tggtcatttc accacgatga tttgactgag agccaacatc gttggatagc
     4321 tccgctggtg gttcatcact ggcccgacta tcaggtacgc tttcccacac gccgtcgtaa
     4381 gctgaacagc ggctactttt gtattacttc tcagcgtttc gctgaggttt tacagcgaca
     4441 gtttggcccg cacttgtgga tggataccgc ggtcgcagag gttaatgcgg aatctgttcg
     4501 gttgaaaaag ggtcaggtta tcggtgcccg cgcggtgatt gacgggcggg gttatgcggc
     4561 aaattcagca ctgagcgtgg gcttccaggc gtttattggc caggaatggc gattgagtca
     4621 cccgcatggt ttatcatctc cgattatcat ggatgccacg gtcgatcagc aaaatggtta
     4681 tcgcttcgtg tacagcctgc cgctctcgcc gaccagattg ttaattgaag atacgcacta
     4741 tattgataat gcgacattag atcctgaatg cgcgcggcaa aatatttgcg actatgccgc
     4801 gcaacagggt tggcagcttc agacactgct gcgagaagaa cagggcgcct tacccattac
     4861 tctgtcgggc aatgccgacg cattctggca gcagcgcccc ctggcctgta gtggattacg
     4921 tgccggtctg ttccatccta ccaccggcta ttcactgccg ctggcggttg ccgtggccga
     4981 ccgcctgagt gcacttgatg tctttacgtc ggcctcaatt caccatgcca ttacgcattt
     5041 tgcccgcgag cgctggcagc agcagggctt tttccgcatg ctgaatcgca tgctgttttt
     5101 agccggaccc gccgattcac gctggcgggt tatgcagcgt ttttatggtt tacctgaaga
     5161 tttaattgcc cgtttttatg cgggaaaact cacgctgacc gatcggctac gtattctgag
     5221 cggcaagccg cctgttccgg tattagcagc attgcaagcc attatgacga ctcatcgtta
     5281 agcttcagag gaggaattaa gtaatgagtt ttgatattgc caaatacccg accctggcac
     5341 tggtcgactc cacccaggag ttacgactgt tgccgaaaga gagtttaccg aaactctgcg
     5401 acgaactgcg ccgctattta ctcgacagcg tgagccgttc cagcgggcac ttcgcctccg
     5461 ggctgggcac ggtcgaactg accgtggcgc tgcactatgt ctacaacacc ccgtttgacc
     5521 aattgatttg ggatgtgggg catcaggctt atccgcataa aattttgacc ggacgccgcg
     5581 acaaaatcgg caccatccgt cagaaaggcg gtctgcaccc gttcccgtgg cgcggcgaaa
     5641 gcgaatatga cgtattaagc gtcgggcatt catcaacctc cattagtgcc ggaattggta
     5701 ttgcggttgc tgccgaaaaa gaaggcaaaa atcgccgcac cgtctgtgtc attggcgatg
     5761 gcgcgattac cgcaggcatg gcgtttgaag cgatgaatca cgcgggcgat atccgtcctg
     5821 atatgctggt gattctcaac gacaatgaaa tgtcgatttc cgaaaatgtc ggcgcgctca
     5881 acaaccatct ggcacagctg ctttccggta agctttactc ttcactgcgc gaaggcggga
     5941 aaaaagtttt ctctggcgtg ccgccaatta aagagctgct caaacgcacc gaagaacata
     6001 ttaaaggcat ggtagtgcct ggcacgttgt ttgaggagct gggctttaac tacatcggcc
     6061 cggtggacgg tcatgatgtg ctggggctta tcaccacgct aaagaacatg cgcgacctga
     6121 aaggcccgca gttcctgcat atcatgacca aaaaaggtcg tggttatgaa ccggcagaaa
     6181 aagacccgat cactttccat gccgtgctaa atttgatccc tccagcggtt gtttgccgaa
     6241 aagtagcggc ggtttgccga gctattcaaa aatctttggc gactggttgt gcgaaacggc
     6301 agcgaaagac aacaagctga tggcgattac tccggcgatg cgtgaaggtt ccggcatggt
     6361 cgagttttca cgtaaattcc cggatcgcta cttcgacgtg gcaattgccg agcaacatgc
     6421 ggtgaccttt gctgcgggtc tggcgattgg tgggtacaaa cccattgtcg cgatttactc
     6481 cactttcctg caacgcgcct atgatcaggt gctgcatgac gtggcgattc aaaagcttcc
     6541 ggtcctgttc gccatcgacc gcgcgggcat tgttggtgct gacggtcaaa cccatcaggg
     6601 tgcttttgat ctctcttacc tgcgctgcat accggaaatg gtcattatga ccccgagcga
     6661 tgaaaacgaa tgtcgccaga tgctctatac cggctatcac tataacgatg gcccgtcagc
     6721 ggtgcgctac ccgcgtggca acgcggtcgg cgtggaactg acgccgctgg aaaaactacc
     6781 aattggcaaa ggcattgtga agcgtcgtgg cgagaaactg gcgatcctta actttggtac
     6841 gctgatgcca gaagcggcga aagtcgccaa atcgctgaac gccacgctgg tcgatatgcg
     6901 ttttgtgaaa ccgcttgatg aagcgttaat tctggaaatg gccgccagcc atgaagcgct
     6961 ggtcaccgta gaagaaaacg ccattatggg cggcgcaggc agcggcgtga acgaagtgct
     7021 gatggcccat cgtaaaccag tacccgtgct gaacattggc ctgccggact tctttattcc
     7081 gcaaggaact caggaagaaa tgcgcgccga actcggcctc gatgccgctg gtatggaagc
     7141 caaaatcaag gcctggctgg cataagcttt gtgtgaggag atgtcgaatg agcgcacatg
     7201 ccctgcccaa ggcagacctg accgccacca gcctgatcgt atcgggcggc atcatcgccg
     7261 cttggctggc cctgcatgtg catgcgctgt ggtttctgga cgcagcggcg catcccatcc
     7321 tggcgatcgc aaatttcctg gggctgacct ggctgtcggt cggattgttc atcatcgcgc
     7381 atgacgcgat gcacgggtcg gtggtgccgg ggcgtccgcg cgccaatgcg gcgatgggcc
     7441 agcttgtcct gtggctgtat gccggatttt cgtggcgcaa gatgatcgtc aagcacatgg
     7501 cccatcaccg ccatgccgga accgacgacg accccgattt cgaccatggc ggcccggtcc
     7561 gctggtacgc ccgcttcatc ggcacctatt tcggctggcg cgaggggctg cttctgcccg
     7621 tcatcgtgac ggtctatgcg ctgatccttg gggatcgctg gatgtacgtg gtattctggc
     7681 cgctgccgtc gatcctggcg tcgatccagc tgttcgtgtt cggcacctgg ctgccgcacc
     7741 gccccggcca cgacgcgttc ccggaccgcc acaatgcgcg gtcgtcgcgg atcagcgacc
     7801 ccgtgtcgct gctgacctgt tttcactttg gcggttatca tcacgaacac cacctgcacc
     7861 cgacggtgcc gtggtggcgc ctgcccagca cccgcaccaa gggcgacacc gcataagctt
     7921 gagctgagga gattttgaat gcaaacggaa cacgtcattt tattgaatgc acagggagtt
     7981 cccacgggta cgctggaaaa gtatgccgca cacacggcag acacccgctt acatctcgcg
     8041 ttctccagtt ggctgtttaa tgccaaagga caattattag ttacccgccg cgcactgagc
     8101 aaaaaagcat ggcctggcgt gtggactaac tcggtttgtg ggcacccaca actgggagaa
     8161 agcaacgagg acgcagtgat ccgccgttgc cgttatgagc ttggcgtgga aattacgcct
     8221 cctgaatcta tctatcctga ctttcgctac cgcgccaccg atccgagtgg cattgtggaa
     8281 aatgaagtgt gtccggtatt tgccgcacgc accactagtg cgttacagat caatgatgat
     8341 gaagtgatgg attatcaatg gtgtgattta gcagatgtat tacacggtat tgatgcaacg
     8401 ccgtgggcgt tcagtccgtg gatggtgatg caggcgacaa atcgcgaagc cagaaaacga
     8461 ttatctgcat ttacccagct taaataagct ttgcctcagg atttttgaaa tgacggtctg
     8521 cgcaaaaaaa cacgttcatc tcactcgcga tgctgcggag cagttactgg ctgatattga
     8581 tcgacgcctt gatcagttat tgcccgtgga gggagaacgg gatgttgtgg gtgccgcgat
     8641 gcgtgaaggt gcgctggcac cgggaaaacg tattcgcccc atgttgctgt tgctgaccgc
     8701 ccgcgatctg ggttgcgctt tcagccatga cggattactg gatttggcct gtgcggtgga
     8761 aatggtccac gcggcttcgc tgatccttga cgatatgccc tgcatggacg atgcgaagct
     8821 gcggcgcgga cgccctacca ttcattctca ttacggagag catgtggcaa tactggcggc
     8881 ggttgccttg ctgagtaaag cctttggcgt aattgccgat gcagatggcc tcacgccgct
     8941 ggcaaaaaat cgggcggttt ctgaactgtc aaacgccatc ggcatgcaag gattggttca
     9001 gggtcagttc aaggatctgt ctgaagggga taagccgcgc agcgctgaag ctattttgat
     9061 gacgaatcac tttaaaacca gcacgctgtt ttgtgcctcg atgcagatgg cctcgattgt
     9121 tgcgaatgcc tccagcgaag cgcgtgattg cctgcatcgt ttttcacttg atcttggtca
     9181 ggcatttcaa ctgctggacg atttgaccga tggcatgacc gacaccggta aggatagcaa
     9241 tcaggacgcc ggtaaatcga cgctggtcaa tctgttaggc ccgagggcgg ttgaagaacg
     9301 tctgagacaa catcttcagc ttgccagtga gcatctctct gcggcctgcc aacacgggca
     9361 cgccactcaa cattttattc aggcctggtt tgacaaaaaa ctcgctgccg tcagttaagc
     9421 ttgcaagact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttggtga
     9481 cggtctcatc gatgagacca agtctccggc aaaaaagggc aaggtgtcac caccctgccc
     9541 tttttcttta aaaccgaaaa gattacttcg cgttatgcag gcttcctcgc tcactgactc
     9601 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     9661 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     9721 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cacaggctcc gcccccctga
     9781 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     9841 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     9901 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     9961 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
    10021 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
    10081 aagacacgac ttatcg
//
