LOCUS       sequence-42952-e        5737 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 5737)
  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 5737)
  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: pBbS5k-RFP
FEATURES             Location/Qualifiers
     source          1..5737
                     /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"
     promoter        join(1520..1525,1526..1543,1544..1550)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1558..1574
                     /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             1615..2292
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2325..2396
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2412..2439
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2970..3920)
                     /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(4682..4776)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4807..5601)
                     /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(4918..4937)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5528..5547
                     /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 tcgtttaggc accccaggct ttacacttta tgcttccggc tcgtataatg tgtggaattg
     1561 tgagcggata acaatttcag aattcaaaag atcttttaag aaggagatat acatatggcg
     1621 agtagcgaag acgttatcaa agagttcatg cgtttcaaag ttcgtatgga aggttccgtt
     1681 aacggtcacg agttcgaaat cgaaggtgaa ggtgaaggtc gtccgtacga aggtacccag
     1741 accgctaaac tgaaagttac caaaggtggt ccgctgccgt tcgcttggga catcctgtcc
     1801 ccgcagttcc agtacggttc caaagcttac gttaaacacc cggctgacat cccggactac
     1861 ctgaaactgt ccttcccgga aggtttcaaa tgggaacgtg ttatgaactt cgaagacggt
     1921 ggtgttgtta ccgttaccca ggactcctcc ctgcaagacg gtgagttcat ctacaaagtt
     1981 aaactgcgtg gtaccaactt cccgtccgac ggtccggtta tgcagaaaaa aaccatgggt
     2041 tgggaagctt ccaccgaacg tatgtacccg gaagacggtg ctctgaaagg tgaaatcaaa
     2101 atgcgtctga aactgaaaga cggtggtcac tacgacgctg aagttaaaac cacctacatg
     2161 gctaaaaaac cggttcagct gccgggtgct tacaaaaccg acatcaaact ggacatcacc
     2221 tcccacaacg aagactacac catcgttgaa cagtacgaac gtgctgaagg tcgtcactcc
     2281 accggtgctt aaggatccaa actcgagtaa ggatctccag gcatcaaata aaacgaaagg
     2341 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact
     2401 agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttatacctag ggtacgggtt
     2461 ttgctgcccg caaacgggct gttctggtgt tgctagtttg ttatcagaat cgcagatccg
     2521 gcttcagccg gtttgccggc tgaaagcgct atttcttcca gaattgccat gattttttcc
     2581 ccacgggagg cgtcactggc tcccgtgttg tcggcagctt tgattcgata agcagcatcg
     2641 cctgtttcag gctgtctatg tgtgactgtt gagctgtaac aagttgtctc aggtgttcaa
     2701 tttcatgttc tagttgcttt gttttactgg tttcacctgt tctattaggt gttacatgct
     2761 gttcatctgt tacattgtcg atctgttcat ggtgaacagc tttgaatgca ccaaaaactc
     2821 gtaaaagctc tgatgtatct atctttttta caccgttttc atctgtgcat atggacagtt
     2881 ttccctttga tatgtaacgg tgaacagttg ttctactttt gtttgttagt cttgatgctt
     2941 cactgataga tacaagagcc ataagaacct cagatccttc cgtatttagc cagtatgttc
     3001 tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg aaccattgag atcatactta
     3061 ctttgcatgt cactcaaaaa ttttgcctca aaactggtga gctgaatttt tgcagttaaa
     3121 gcatcgtgta gtgtttttct tagtccgtta tgtaggtagg aatctgatgt aatggttgtt
     3181 ggtattttgt caccattcat ttttatctgg ttgttctcaa gttcggttac gagatccatt
     3241 tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg ggcggcctcg cttatcaacc
     3301 accaatttca tattgctgta agtgtttaaa tctttactta ttggtttcaa aacccattgg
     3361 ttaagccttt taaactcatg gtagttattt tcaagcatta acatgaactt aaattcatca
     3421 aggctaatct ctatatttgc cttgtgagtt ttcttttgtg ttagttcttt taataaccac
     3481 tcataaatcc tcatagagta tttgttttca aaagacttaa catgttccag attatatttt
     3541 atgaattttt ttaactggaa aagataaggc aatatctctt cactaaaaac taattctaat
     3601 ttttcgcttg agaacttggc atagtttgtc cactggaaaa tctcaaagcc tttaaccaaa
     3661 ggattcctga tttccacagt tctcgtcatc agctctctgg ttgctttagc taatacacca
     3721 taagcatttt ccctactgat gttcatcatc tgagcgtatt ggttataagt gaacgatacc
     3781 gtccgttctt tccttgtagg gttttcaatc gtggggttga gtagtgccac acagcataaa
     3841 attagcttgg tttcatgctc cgttaagtca tagcgactaa tcgctagttc atttgctttg
     3901 aaaacaacta attcagacat acatctcaat tggtctaggt gattttaatc actataccaa
     3961 ttgagatggg ctagtcaatg ataattacta gtccttttcc cgggtgatct gggtatctgt
     4021 aaattctgct agacctttgc tggaaaactt gtaaattctg ctagaccctc tgtaaattcc
     4081 gctagacctt tgtgtgtttt ttttgtttat attcaagtgg ttataattta tagaataaag
     4141 aaagaataaa aaaagataaa aagaatagat cccagccctg tgtataactc actactttag
     4201 tcagttccgc agtattacaa aaggatgtcg caaacgctgt ttgctcctct acaaaacaga
     4261 ccttaaaacc ctaaaggctt aagtagcacc ctcgcaagct cgggcaaatc gctgaatatt
     4321 ccttttgtct ccgaccatca ggcacctgag tcgctgtctt tttcgtgaca ttcagttcgc
     4381 tgcgctcacg gctctggcag tgaatggggg taaatggcac tacaggcgcc ttttatggat
     4441 tcatgcaagg aaactaccca taatacaaga aaagcccgtc acgggcttct cagggcgttt
     4501 tatggcgggt ctgctatgtg gtgctatctg actttttgct gttcagcagt tcctgccctc
     4561 tgattttcca gtctgaccac ttcggattat cccgtgacag gtcattcaga ctggctaatg
     4621 cacccagtaa ggcagcggta tcatcaacag gcttacccgt cttactgtcc ctagtgcttg
     4681 gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga
     4741 gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctctcg aaccccagag
     4801 tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc
     4861 ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat
     4921 atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc
     4981 gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg
     5041 ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg agcctggcga acagttcggc
     5101 tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat
     5161 ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg
     5221 atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc
     5281 aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc
     5341 cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga
     5401 tagccgcgct gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa
     5461 aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt
     5521 ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg
     5581 caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cagatcatga
     5641 tcccctgcgc catcagatcc ttggcggcaa gaaagccatc cagtttactt tgcagggctt
     5701 cccaacctta ccagagggcg ccccagctgg caattcc
//
