LOCUS       sequence-42988-e        5683 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 5683)
  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 5683)
  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: pBbS7k-RFP
FEATURES             Location/Qualifiers
     source          1..5683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(8..1090)
                     /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     1052..1071
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1091..1168)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1374..1393
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1481..1500
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1481..1499
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1504..1520
                     /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             1561..2238
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2271..2342
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2358..2385
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2916..3866)
                     /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(4628..4722)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4753..5547)
                     /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(4864..4883)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5474..5493
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
       61 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
      121 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
      181 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
      241 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg tccgcaccaa
      301 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
      361 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
      421 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
      481 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
      541 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
      601 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
      661 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
      721 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
      781 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
      841 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
      901 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
      961 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     1021 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
     1081 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     1141 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
     1201 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     1261 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     1321 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     1381 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     1441 ccggcgtaga ggatcgagat cgatctcgat cccgcgaaat taatacgact cactataggg
     1501 gaattgtgag cggataacaa tttcagaatt caaaagatct tttaagaagg agatatacat
     1561 atggcgagta gcgaagacgt tatcaaagag ttcatgcgtt tcaaagttcg tatggaaggt
     1621 tccgttaacg gtcacgagtt cgaaatcgaa ggtgaaggtg aaggtcgtcc gtacgaaggt
     1681 acccagaccg ctaaactgaa agttaccaaa ggtggtccgc tgccgttcgc ttgggacatc
     1741 ctgtccccgc agttccagta cggttccaaa gcttacgtta aacacccggc tgacatcccg
     1801 gactacctga aactgtcctt cccggaaggt ttcaaatggg aacgtgttat gaacttcgaa
     1861 gacggtggtg ttgttaccgt tacccaggac tcctccctgc aagacggtga gttcatctac
     1921 aaagttaaac tgcgtggtac caacttcccg tccgacggtc cggttatgca gaaaaaaacc
     1981 atgggttggg aagcttccac cgaacgtatg tacccggaag acggtgctct gaaaggtgaa
     2041 atcaaaatgc gtctgaaact gaaagacggt ggtcactacg acgctgaagt taaaaccacc
     2101 tacatggcta aaaaaccggt tcagctgccg ggtgcttaca aaaccgacat caaactggac
     2161 atcacctccc acaacgaaga ctacaccatc gttgaacagt acgaacgtgc tgaaggtcgt
     2221 cactccaccg gtgcttaagg atccaaactc gagtaaggat ctccaggcat caaataaaac
     2281 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     2341 tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat acctagggta
     2401 cgggttttgc tgcccgcaaa cgggctgttc tggtgttgct agtttgttat cagaatcgca
     2461 gatccggctt cagccggttt gccggctgaa agcgctattt cttccagaat tgccatgatt
     2521 ttttccccac gggaggcgtc actggctccc gtgttgtcgg cagctttgat tcgataagca
     2581 gcatcgcctg tttcaggctg tctatgtgtg actgttgagc tgtaacaagt tgtctcaggt
     2641 gttcaatttc atgttctagt tgctttgttt tactggtttc acctgttcta ttaggtgtta
     2701 catgctgttc atctgttaca ttgtcgatct gttcatggtg aacagctttg aatgcaccaa
     2761 aaactcgtaa aagctctgat gtatctatct tttttacacc gttttcatct gtgcatatgg
     2821 acagttttcc ctttgatatg taacggtgaa cagttgttct acttttgttt gttagtcttg
     2881 atgcttcact gatagataca agagccataa gaacctcaga tccttccgta tttagccagt
     2941 atgttctcta gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc attgagatca
     3001 tacttacttt gcatgtcact caaaaatttt gcctcaaaac tggtgagctg aatttttgca
     3061 gttaaagcat cgtgtagtgt ttttcttagt ccgttatgta ggtaggaatc tgatgtaatg
     3121 gttgttggta ttttgtcacc attcattttt atctggttgt tctcaagttc ggttacgaga
     3181 tccatttgtc tatctagttc aacttggaaa atcaacgtat cagtcgggcg gcctcgctta
     3241 tcaaccacca atttcatatt gctgtaagtg tttaaatctt tacttattgg tttcaaaacc
     3301 cattggttaa gccttttaaa ctcatggtag ttattttcaa gcattaacat gaacttaaat
     3361 tcatcaaggc taatctctat atttgccttg tgagttttct tttgtgttag ttcttttaat
     3421 aaccactcat aaatcctcat agagtatttg ttttcaaaag acttaacatg ttccagatta
     3481 tattttatga atttttttaa ctggaaaaga taaggcaata tctcttcact aaaaactaat
     3541 tctaattttt cgcttgagaa cttggcatag tttgtccact ggaaaatctc aaagccttta
     3601 accaaaggat tcctgatttc cacagttctc gtcatcagct ctctggttgc tttagctaat
     3661 acaccataag cattttccct actgatgttc atcatctgag cgtattggtt ataagtgaac
     3721 gataccgtcc gttctttcct tgtagggttt tcaatcgtgg ggttgagtag tgccacacag
     3781 cataaaatta gcttggtttc atgctccgtt aagtcatagc gactaatcgc tagttcattt
     3841 gctttgaaaa caactaattc agacatacat ctcaattggt ctaggtgatt ttaatcacta
     3901 taccaattga gatgggctag tcaatgataa ttactagtcc ttttcccggg tgatctgggt
     3961 atctgtaaat tctgctagac ctttgctgga aaacttgtaa attctgctag accctctgta
     4021 aattccgcta gacctttgtg tgtttttttt gtttatattc aagtggttat aatttataga
     4081 ataaagaaag aataaaaaaa gataaaaaga atagatccca gccctgtgta taactcacta
     4141 ctttagtcag ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa
     4201 aacagacctt aaaaccctaa aggcttaagt agcaccctcg caagctcggg caaatcgctg
     4261 aatattcctt ttgtctccga ccatcaggca cctgagtcgc tgtctttttc gtgacattca
     4321 gttcgctgcg ctcacggctc tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt
     4381 atggattcat gcaaggaaac tacccataat acaagaaaag cccgtcacgg gcttctcagg
     4441 gcgttttatg gcgggtctgc tatgtggtgc tatctgactt tttgctgttc agcagttcct
     4501 gccctctgat tttccagtct gaccacttcg gattatcccg tgacaggtca ttcagactgg
     4561 ctaatgcacc cagtaaggca gcggtatcat caacaggctt acccgtctta ctgtccctag
     4621 tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca
     4681 gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag ctctcgaacc
     4741 ccagagtccc gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc
     4801 gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc
     4861 agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc
     4921 acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc
     4981 gccatgggtc acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag
     5041 ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc
     5101 ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt
     5161 agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc
     5221 aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc
     5281 ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag
     5341 ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt
     5401 gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc
     5461 gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc
     5521 tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcaga
     5581 tcatgatccc ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca
     5641 gggcttccca accttaccag agggcgcccc agctggcaat tcc
//
