LOCUS       sequence-42961-e        5731 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 5731)
  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 5731)
  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: pBbS6k-RFP
FEATURES             Location/Qualifiers
     source          1..5731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(49..1131)
                     /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     1093..1112
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1132..1209)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1415..1434
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     protein_bind    1502..1518
                     /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)."
     protein_bind    1525..1541
                     /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             1609..2286
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2319..2390
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2406..2433
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2964..3914)
                     /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(4676..4770)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4801..5595)
                     /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(4912..4931)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5522..5541
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
       61 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      121 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
      181 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
      241 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
      301 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt
      361 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
      421 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
      481 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
      541 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
      601 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
      661 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
      721 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
      781 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
      841 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
      901 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
      961 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     1021 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     1081 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     1141 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     1201 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     1261 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     1321 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     1381 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     1441 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     1501 attgtgagcg gataacaatt gacattgtga gcggataaca agatactgag cacatcagca
     1561 ggacgcactg accgaattca aaagatcttt taagaaggag atatacatat ggcgagtagc
     1621 gaagacgtta tcaaagagtt catgcgtttc aaagttcgta tggaaggttc cgttaacggt
     1681 cacgagttcg aaatcgaagg tgaaggtgaa ggtcgtccgt acgaaggtac ccagaccgct
     1741 aaactgaaag ttaccaaagg tggtccgctg ccgttcgctt gggacatcct gtccccgcag
     1801 ttccagtacg gttccaaagc ttacgttaaa cacccggctg acatcccgga ctacctgaaa
     1861 ctgtccttcc cggaaggttt caaatgggaa cgtgttatga acttcgaaga cggtggtgtt
     1921 gttaccgtta cccaggactc ctccctgcaa gacggtgagt tcatctacaa agttaaactg
     1981 cgtggtacca acttcccgtc cgacggtccg gttatgcaga aaaaaaccat gggttgggaa
     2041 gcttccaccg aacgtatgta cccggaagac ggtgctctga aaggtgaaat caaaatgcgt
     2101 ctgaaactga aagacggtgg tcactacgac gctgaagtta aaaccaccta catggctaaa
     2161 aaaccggttc agctgccggg tgcttacaaa accgacatca aactggacat cacctcccac
     2221 aacgaagact acaccatcgt tgaacagtac gaacgtgctg aaggtcgtca ctccaccggt
     2281 gcttaaggat ccaaactcga gtaaggatct ccaggcatca aataaaacga aaggctcagt
     2341 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc
     2401 acactggctc accttcgggt gggcctttct gcgtttatac ctagggtacg ggttttgctg
     2461 cccgcaaacg ggctgttctg gtgttgctag tttgttatca gaatcgcaga tccggcttca
     2521 gccggtttgc cggctgaaag cgctatttct tccagaattg ccatgatttt ttccccacgg
     2581 gaggcgtcac tggctcccgt gttgtcggca gctttgattc gataagcagc atcgcctgtt
     2641 tcaggctgtc tatgtgtgac tgttgagctg taacaagttg tctcaggtgt tcaatttcat
     2701 gttctagttg ctttgtttta ctggtttcac ctgttctatt aggtgttaca tgctgttcat
     2761 ctgttacatt gtcgatctgt tcatggtgaa cagctttgaa tgcaccaaaa actcgtaaaa
     2821 gctctgatgt atctatcttt tttacaccgt tttcatctgt gcatatggac agttttccct
     2881 ttgatatgta acggtgaaca gttgttctac ttttgtttgt tagtcttgat gcttcactga
     2941 tagatacaag agccataaga acctcagatc cttccgtatt tagccagtat gttctctagt
     3001 gtggttcgtt gtttttgcgt gagccatgag aacgaaccat tgagatcata cttactttgc
     3061 atgtcactca aaaattttgc ctcaaaactg gtgagctgaa tttttgcagt taaagcatcg
     3121 tgtagtgttt ttcttagtcc gttatgtagg taggaatctg atgtaatggt tgttggtatt
     3181 ttgtcaccat tcatttttat ctggttgttc tcaagttcgg ttacgagatc catttgtcta
     3241 tctagttcaa cttggaaaat caacgtatca gtcgggcggc ctcgcttatc aaccaccaat
     3301 ttcatattgc tgtaagtgtt taaatcttta cttattggtt tcaaaaccca ttggttaagc
     3361 cttttaaact catggtagtt attttcaagc attaacatga acttaaattc atcaaggcta
     3421 atctctatat ttgccttgtg agttttcttt tgtgttagtt cttttaataa ccactcataa
     3481 atcctcatag agtatttgtt ttcaaaagac ttaacatgtt ccagattata ttttatgaat
     3541 ttttttaact ggaaaagata aggcaatatc tcttcactaa aaactaattc taatttttcg
     3601 cttgagaact tggcatagtt tgtccactgg aaaatctcaa agcctttaac caaaggattc
     3661 ctgatttcca cagttctcgt catcagctct ctggttgctt tagctaatac accataagca
     3721 ttttccctac tgatgttcat catctgagcg tattggttat aagtgaacga taccgtccgt
     3781 tctttccttg tagggttttc aatcgtgggg ttgagtagtg ccacacagca taaaattagc
     3841 ttggtttcat gctccgttaa gtcatagcga ctaatcgcta gttcatttgc tttgaaaaca
     3901 actaattcag acatacatct caattggtct aggtgatttt aatcactata ccaattgaga
     3961 tgggctagtc aatgataatt actagtcctt ttcccgggtg atctgggtat ctgtaaattc
     4021 tgctagacct ttgctggaaa acttgtaaat tctgctagac cctctgtaaa ttccgctaga
     4081 cctttgtgtg ttttttttgt ttatattcaa gtggttataa tttatagaat aaagaaagaa
     4141 taaaaaaaga taaaaagaat agatcccagc cctgtgtata actcactact ttagtcagtt
     4201 ccgcagtatt acaaaaggat gtcgcaaacg ctgtttgctc ctctacaaaa cagaccttaa
     4261 aaccctaaag gcttaagtag caccctcgca agctcgggca aatcgctgaa tattcctttt
     4321 gtctccgacc atcaggcacc tgagtcgctg tctttttcgt gacattcagt tcgctgcgct
     4381 cacggctctg gcagtgaatg ggggtaaatg gcactacagg cgccttttat ggattcatgc
     4441 aaggaaacta cccataatac aagaaaagcc cgtcacgggc ttctcagggc gttttatggc
     4501 gggtctgcta tgtggtgcta tctgactttt tgctgttcag cagttcctgc cctctgattt
     4561 tccagtctga ccacttcgga ttatcccgtg acaggtcatt cagactggct aatgcaccca
     4621 gtaaggcagc ggtatcatca acaggcttac ccgtcttact gtccctagtg cttggattct
     4681 caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg
     4741 aggtcattac tggatctatc aacaggagtc caagcgagct ctcgaacccc agagtcccgc
     4801 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
     4861 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
     4921 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
     4981 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
     5041 gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc
     5101 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
     5161 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
     5221 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
     5281 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
     5341 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
     5401 cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
     5461 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
     5521 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
     5581 atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc atgatcccct
     5641 gcgccatcag atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac
     5701 cttaccagag ggcgccccag ctggcaattc c
//
