LOCUS       sequence-42949-e        4315 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 4315)
  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 4315)
  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: pBbA5k-RFP
FEATURES             Location/Qualifiers
     source          1..4315
                     /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"
     primer_bind     complement(2467..2484)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2601..3146)
                     /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(2681..2700)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3260..3354)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3385..4179)
                     /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(3496..3515)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4106..4125
                     /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 ggatatattc
     2461 cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc
     2521 ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag
     2581 agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc
     2641 tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2701 cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg
     2761 ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg
     2821 ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg
     2881 taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac
     2941 tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa
     3001 ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag
     3061 ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga
     3121 ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata aaatatttct
     3181 agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata
     3241 agttgttact agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc
     3301 tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg
     3361 agctctcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
     3421 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
     3481 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
     3541 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
     3601 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag
     3661 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
     3721 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
     3781 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     3841 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     3901 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     3961 cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga
     4021 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     4081 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     4141 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
     4201 ctcttgatca gatcatgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca
     4261 gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca attcc
//
