LOCUS       sequence-455391-        4675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BirA encoded pACYC vector for co-expression with Avi-tagged 
            constructs (Bacterial Expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4675)
  TITLE     Molecular Toolbox
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4675)
  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            "Unpublished"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: MZB209
FEATURES             Location/Qualifiers
     source          1..4675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(188..847)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     762..781
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(848..950)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(1342..1359)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1476..2021)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1556..1575)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2246..3328)
                     /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     3290..3309
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3329..3406)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3477..3496
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3477..3495
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3527..3549
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3556..4521
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             3556..4518
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     primer_bind     complement(4581..4599)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4595..4642
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 acactgcttc cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa
       61 cgaccctgcc ctgaaccgac gaccgggtcg aatttgcttt cgaatttctg ccattcatcc
      121 gcttattatc acttattcag gcgtagcaac caggcgttta agggcaccaa taactgcctt
      181 aaaaaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
      241 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
      301 ccttgtcgcc ttgcgtataa tatttgccca tagtgaaaac gggggcgaag aagttgtcca
      361 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa
      421 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat
      481 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg
      541 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
      601 ccagctcacc gtctttcatt gccatacgga actccggatg agcattcatc aggcgggcaa
      661 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
      721 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
      781 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
      841 tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
      901 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt
      961 ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg gacaccagga tttatttatt
     1021 ctgcgaagtg atcttccgtc acaggtattt attcggcgca aagtgcgtcg ggtgatgctg
     1081 ccaacttact gatttagtgt atgatggtgt ttttgaggtg ctccagtggc ttctgtttct
     1141 atcagctgtc cctcctgttc agctactgac ggggtggtgc gtaacggcaa aagcaccgcc
     1201 ggacatcagc gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca
     1261 gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg
     1321 atacaggata tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg
     1381 gcgagcggaa atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact
     1441 taacagggaa gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac
     1501 aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga
     1561 taccaggcgt ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt
     1621 accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc
     1681 cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg
     1741 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc
     1801 actggcagca gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt
     1861 aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt
     1921 tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt
     1981 tttcagagca agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc
     2041 agataaaata tttctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca
     2101 gccccatacg atataagttg taattctcat gttagtcatg ccccgcgccc accggaagga
     2161 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
     2221 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2281 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     2341 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
     2401 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
     2461 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
     2521 ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
     2581 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     2641 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     2701 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
     2761 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
     2821 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
     2881 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     2941 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     3001 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     3061 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     3121 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     3181 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     3241 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     3301 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     3361 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc
     3421 tctcccttat gcgactcctg cattaggcgc gaggcaggat ctcgatcccg cgaaattaat
     3481 acgactcact atagggagac cacaacggtt tccctctaga aataattttg tttaacttta
     3541 agaaggagat ataccatgaa ggataacacc gtgccactga aattgattgc cctgttagcg
     3601 aacggtgaat ttcactctgg cgagcagttg ggtgaaacgc tgggaatgag ccgggcggct
     3661 attaataaac acattcagac actgcgtgac tggggcgttg atgtctttac cgttccgggt
     3721 aaaggataca gcctgcctga gcctatccag ttacttaatg ctaaacagat attgggtcag
     3781 ctggatggcg gtagtgtagc cgtgctgcct gtgattgact ccacgaatca gtaccttctt
     3841 gatcgtatcg gagagcttaa atcgggcgat gcttgcattg cagaatacca gcaggctggc
     3901 cgtggtcgcc ggggtcggaa atggttttcg ccttttggcg caaacttata tttgtcgatg
     3961 ttctggcgtc tggaacaagg cccggcggcg gcgattggtt taagtctggt tatcggtatc
     4021 gtgatggcgg aagtattacg caagctgggt gcagataaag ttcgtgttaa atggcctaat
     4081 gacctctatc tgcaggatcg caagctggca ggcattctgg tggagctgac tggcaaaact
     4141 ggcgatgcgg cgcaaatagt cattggagcc gggatcaaca tggcaatgcg ccgtgttgaa
     4201 gagagtgtcg ttaatcaggg gtggatcacg ctgcaggaag cggggatcaa tctcgatcgt
     4261 aatacgttgg cggccatgct aatacgtgaa ttacgtgctg cgttggaact cttcgaacaa
     4321 gaaggattgg caccttatct gtcgcgctgg gaaaagctgg ataattttat taatcgccca
     4381 gtgaaactta tcattggtga taaagaaata tttggcattt cacgcggaat agacaaacag
     4441 ggggctttat tacttgagca ggatggaata ataaaaccct ggatgggcgg tgaaatatcc
     4501 ctgcgtagtg cagaaaaata ataagaattc cgatccggct gctaacaaag cccgaaagga
     4561 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     4621 acgggtcttg aggggttttt tgctgaaacc tcaggcattt gagaagcaca cggtc
//
