LOCUS       sequence-42985-e        4261 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 4261)
  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 4261)
  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: pBbA7k-RFP
FEATURES             Location/Qualifiers
     source          1..4261
                     /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"
     primer_bind     complement(2413..2430)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2547..3092)
                     /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(2627..2646)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3206..3300)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3331..4125)
                     /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(3442..3461)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4052..4071
                     /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 acctagggat
     2401 atattccgct tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga
     2461 aatggcttac gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga
     2521 agtgagaggg ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac
     2581 gaaatctgac gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg
     2641 tttccccctg gcggctccct cgtgcgctct cctgttcctg cctttcggtt taccggtgtc
     2701 attccgctgt tatggccgcg tttgtctcat tccacgcctg acactcagtt ccgggtaggc
     2761 agttcgctcc aagctggact gtatgcacga accccccgtt cagtccgacc gctgcgcctt
     2821 atccggtaac tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac cactggcagc
     2881 agccactggt aattgattta gaggagttag tcttgaagtc atgcgccggt taaggctaaa
     2941 ctgaaaggac aagttttggt gactgcgctc ctccaagcca gttacctcgg ttcaaagagt
     3001 tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc
     3061 aagagattac gcgcagacca aaacgatctc aagaagatca tcttattaat cagataaaat
     3121 atttctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac
     3181 gatataagtt gttactagtg cttggattct caccaataaa aaacgcccgg cggcaaccga
     3241 gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc
     3301 caagcgagct ctcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa
     3361 ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca
     3421 ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc
     3481 cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat
     3541 attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc
     3601 cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc
     3661 ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg
     3721 gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat
     3781 gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc
     3841 gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg
     3901 aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt cattcagggc
     3961 accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac
     4021 ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac
     4081 ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca
     4141 tcctgtctct tgatcagatc atgatcccct gcgccatcag atccttggcg gcaagaaagc
     4201 catccagttt actttgcagg gcttcccaac cttaccagag ggcgccccag ctggcaattc
     4261 c
//
