LOCUS       sequence-471455-        2172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gene10 5'UTR T7 3a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2172)
  AUTHORS   Inckemann R, Chotel T, Brinkmann CK, Burgis M, Andreas L, Baumann J,
            Sharma P, Klose M, Barrett J, Ries F, Paczia N, Glatter T, Willmund 
            F, Mackinder LCM, Erb TJ
  TITLE     Advancing chloroplast synthetic biology through high-throughput 
            plastome engineering of Chlamydomonas reinhardtii
  JOURNAL   bioRxiv 2024.05.08.593163
REFERENCE   2  (bases 1 to 2172)
  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: "bioRxiv 
            2024.05.08.593163"
COMMENT     Sequence Label: pME_Cp_0_3a_001_gene10_5'UTR_T7
FEATURES             Location/Qualifiers
     source          1..2172
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      209..797
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     698..717
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     951..968
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(992..1049)
                     /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."
     RBS             1098..1120
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    1173..1194
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      1197..1240
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(1244..1262)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             1424..2083
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1490..1509)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      join(2104..2172,1..26)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 gccgggcgtt ttttattggt gagaatccaa gcctcgagct gtcagaccaa gtttactcat
       61 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
      121 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
      181 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
      241 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
      301 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
      361 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
      421 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
      481 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
      541 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
      601 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
      661 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
      721 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
      781 ggcggagcct gtggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
      841 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
      901 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
      961 tgagcgagga agcctgcata acgcgaagta atcttttcgg ttttaaagaa aaagggcagg
     1021 gtggtgacac cttgcccttt tttgccggac tgcagcggcc gctactagtc gcgatgagtg
     1081 gtctcaatgg atgtatatct ccttcttaaa gttaaacaaa attatttcta gtgggaaacc
     1141 gttgtggtct gccagtacga gaccactcat cgctctagaa gcggccgcga attccagaaa
     1201 tcatccttag cgaaagctaa ggattttttt tatctgaaat tctgcctcgt gatacgccta
     1261 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cactcgagtt
     1321 gatcgggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt
     1381 attttttgag ttatcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac
     1441 tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca
     1501 gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa
     1561 gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct
     1621 gatgaatgct catccggaat ttcgtatggc aatgaaagac ggtgagctgg tgatatggga
     1681 tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcactctg
     1741 gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg
     1801 ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtatc
     1861 agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt
     1921 cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc
     1981 gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa
     2041 tgaattacaa cagtactgcg acgagtggca gggcggggcg taatttgata tcgagctcgc
     2101 ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa
     2161 cgctcggttg cc
//
