LOCUS       sequence-43006-e        5615 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5615)
  AUTHORS   Lee TS, Krupa RA, Zhang F, Hajimorad M, Holtz WJ, Prasad N, Lee SK, 
            Keasling JD
  TITLE     BglBrick vectors and datasheets: A synthetic biology platform for 
            gene expression.
  JOURNAL   J Biol Eng. 2011 Sep 20;5:12.
  PUBMED    21933410
REFERENCE   2  (bases 1 to 5615)
  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: "J Biol 
            Eng."; date: "2011-09-20"; volume: "5"; pages: "12"
COMMENT     Sequence Label: pBbS1k-RFP
FEATURES             Location/Qualifiers
     source          1..5615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..84
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             85..1167
                     /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     complement(104..123)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1399..1404,1405..1421,1422..1428)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1436..1452
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by 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)."
     CDS             1493..2170
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2203..2274
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2290..2317
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2848..3798)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(4560..4654)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4685..5479)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4796..4815)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5406..5425
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcgaca ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa
       61 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
      121 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
      181 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
      241 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
      301 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
      361 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
      421 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
      481 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
      541 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
      601 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
      661 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
      721 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
      781 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
      841 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
      901 tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat
      961 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     1021 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     1081 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     1141 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agcgcgaatt
     1201 gatctggttt gacagcttat catcgactgc acggtgcacc aatgcttctg gcgtcaggca
     1261 gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct
     1321 caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa
     1381 atattctgaa atgagctgtt gacaattaat catccggctc gtataatgtg tggaattgtg
     1441 agcggataac aatttcagaa ttcaaaagat cttttaagaa ggagatatac atatggcgag
     1501 tagcgaagac gttatcaaag agttcatgcg tttcaaagtt cgtatggaag gttccgttaa
     1561 cggtcacgag ttcgaaatcg aaggtgaagg tgaaggtcgt ccgtacgaag gtacccagac
     1621 cgctaaactg aaagttacca aaggtggtcc gctgccgttc gcttgggaca tcctgtcccc
     1681 gcagttccag tacggttcca aagcttacgt taaacacccg gctgacatcc cggactacct
     1741 gaaactgtcc ttcccggaag gtttcaaatg ggaacgtgtt atgaacttcg aagacggtgg
     1801 tgttgttacc gttacccagg actcctccct gcaagacggt gagttcatct acaaagttaa
     1861 actgcgtggt accaacttcc cgtccgacgg tccggttatg cagaaaaaaa ccatgggttg
     1921 ggaagcttcc accgaacgta tgtacccgga agacggtgct ctgaaaggtg aaatcaaaat
     1981 gcgtctgaaa ctgaaagacg gtggtcacta cgacgctgaa gttaaaacca cctacatggc
     2041 taaaaaaccg gttcagctgc cgggtgctta caaaaccgac atcaaactgg acatcacctc
     2101 ccacaacgaa gactacacca tcgttgaaca gtacgaacgt gctgaaggtc gtcactccac
     2161 cggtgcttaa ggatccaaac tcgagtaagg atctccaggc atcaaataaa acgaaaggct
     2221 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag
     2281 agtcacactg gctcaccttc gggtgggcct ttctgcgttt atacctaggg tacgggtttt
     2341 gctgcccgca aacgggctgt tctggtgttg ctagtttgtt atcagaatcg cagatccggc
     2401 ttcagccggt ttgccggctg aaagcgctat ttcttccaga attgccatga ttttttcccc
     2461 acgggaggcg tcactggctc ccgtgttgtc ggcagctttg attcgataag cagcatcgcc
     2521 tgtttcaggc tgtctatgtg tgactgttga gctgtaacaa gttgtctcag gtgttcaatt
     2581 tcatgttcta gttgctttgt tttactggtt tcacctgttc tattaggtgt tacatgctgt
     2641 tcatctgtta cattgtcgat ctgttcatgg tgaacagctt tgaatgcacc aaaaactcgt
     2701 aaaagctctg atgtatctat cttttttaca ccgttttcat ctgtgcatat ggacagtttt
     2761 ccctttgata tgtaacggtg aacagttgtt ctacttttgt ttgttagtct tgatgcttca
     2821 ctgatagata caagagccat aagaacctca gatccttccg tatttagcca gtatgttctc
     2881 tagtgtggtt cgttgttttt gcgtgagcca tgagaacgaa ccattgagat catacttact
     2941 ttgcatgtca ctcaaaaatt ttgcctcaaa actggtgagc tgaatttttg cagttaaagc
     3001 atcgtgtagt gtttttctta gtccgttatg taggtaggaa tctgatgtaa tggttgttgg
     3061 tattttgtca ccattcattt ttatctggtt gttctcaagt tcggttacga gatccatttg
     3121 tctatctagt tcaacttgga aaatcaacgt atcagtcggg cggcctcgct tatcaaccac
     3181 caatttcata ttgctgtaag tgtttaaatc tttacttatt ggtttcaaaa cccattggtt
     3241 aagcctttta aactcatggt agttattttc aagcattaac atgaacttaa attcatcaag
     3301 gctaatctct atatttgcct tgtgagtttt cttttgtgtt agttctttta ataaccactc
     3361 ataaatcctc atagagtatt tgttttcaaa agacttaaca tgttccagat tatattttat
     3421 gaattttttt aactggaaaa gataaggcaa tatctcttca ctaaaaacta attctaattt
     3481 ttcgcttgag aacttggcat agtttgtcca ctggaaaatc tcaaagcctt taaccaaagg
     3541 attcctgatt tccacagttc tcgtcatcag ctctctggtt gctttagcta atacaccata
     3601 agcattttcc ctactgatgt tcatcatctg agcgtattgg ttataagtga acgataccgt
     3661 ccgttctttc cttgtagggt tttcaatcgt ggggttgagt agtgccacac agcataaaat
     3721 tagcttggtt tcatgctccg ttaagtcata gcgactaatc gctagttcat ttgctttgaa
     3781 aacaactaat tcagacatac atctcaattg gtctaggtga ttttaatcac tataccaatt
     3841 gagatgggct agtcaatgat aattactagt ccttttcccg ggtgatctgg gtatctgtaa
     3901 attctgctag acctttgctg gaaaacttgt aaattctgct agaccctctg taaattccgc
     3961 tagacctttg tgtgtttttt ttgtttatat tcaagtggtt ataatttata gaataaagaa
     4021 agaataaaaa aagataaaaa gaatagatcc cagccctgtg tataactcac tactttagtc
     4081 agttccgcag tattacaaaa ggatgtcgca aacgctgttt gctcctctac aaaacagacc
     4141 ttaaaaccct aaaggcttaa gtagcaccct cgcaagctcg ggcaaatcgc tgaatattcc
     4201 ttttgtctcc gaccatcagg cacctgagtc gctgtctttt tcgtgacatt cagttcgctg
     4261 cgctcacggc tctggcagtg aatgggggta aatggcacta caggcgcctt ttatggattc
     4321 atgcaaggaa actacccata atacaagaaa agcccgtcac gggcttctca gggcgtttta
     4381 tggcgggtct gctatgtggt gctatctgac tttttgctgt tcagcagttc ctgccctctg
     4441 attttccagt ctgaccactt cggattatcc cgtgacaggt cattcagact ggctaatgca
     4501 cccagtaagg cagcggtatc atcaacaggc ttacccgtct tactgtccct agtgcttgga
     4561 ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt
     4621 tctgaggtca ttactggatc tatcaacagg agtccaagcg agctctcgaa ccccagagtc
     4681 ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg
     4741 cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat
     4801 cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga
     4861 tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg
     4921 tcacgacgag atcctcgccg tcgggcatgc gcgccttgag cctggcgaac agttcggctg
     4981 gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc
     5041 gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat
     5101 caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa
     5161 ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg
     5221 cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata
     5281 gccgcgctgc ctcgtcctgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa
     5341 gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct
     5401 gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca
     5461 atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatca gatcatgatc
     5521 ccctgcgcca tcagatcctt ggcggcaaga aagccatcca gtttactttg cagggcttcc
     5581 caaccttacc agagggcgcc ccagctggca attcc
//
