LOCUS       sequence-482798-        4789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GUS:NPTII repair template used for HDR experiments with compatible 
            reporter lines. Has B6 Goldenbraid grammar..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4789)
  TITLE     Alonso lab plasmids 2025
REFERENCE   2  (bases 1 to 4789)
  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"
COMMENT     Sequence Label: RT B6-pUPD2
FEATURES             Location/Qualifiers
     source          1..4789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(189..283)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(304..963)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     878..897
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1125..1143
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1147..1190)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1193..1214
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             2739..3530
                     /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="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2790..2809)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3400..3419
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      3557..3809
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3833..3852)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3834..3852)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      3955..4012
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(4036..4053)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4207..4789,1..6))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4287..4306)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
       61 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
      121 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagctcg
      181 aggcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc
      241 agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgatatc
      301 aaattacgcc ccgccctgcc actcgtcgca gtactgttgt aattcattaa gcattctgcc
      361 gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt
      421 gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt
      481 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgata cgaaaaacat
      541 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
      601 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gtgatgaaaa
      661 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag
      721 ctcaccgtct ttcattgcca tacgaaattc cggatgagca ttcatcaggc gggcaagaat
      781 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt
      841 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa
      901 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc
      961 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     1021 tcttatttca ttatggtgaa agttggaacc tcttacgtgc ccgatcaact cgagtgccac
     1081 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     1141 ggcagaattt cagataaaaa aaatccttag ctttcgctaa ggatgatttc tggaattcgc
     1201 ggccgcttct agagcgatga gtggtctcgg cttggcgtgc ctgcagacta gtatttaaat
     1261 tagggataac agggtaatag cagtcttact tccatgattt ctttaactat gccggaatcc
     1321 atcgcagcgt aatgctctac accacgccga acacctgggt ggacgatatc accgtggtga
     1381 cgcatgtcgc gcaagactgt aaccacgcgt ctgttgactg gcaggtggtg gccaatggtg
     1441 atgtcagcgt tgaactgcgt gatgcggatc aacaggtggt tgcaactgga caaggcacta
     1501 gcgggacttt gcaagtggtg aatccgcacc tctggcaacc gggtgaaggt tatctctatg
     1561 aactgtgcgt cacagccaaa agccagacag agtgtgatat ctacccgctt cgcgtcggca
     1621 tccggtcagt ggcagtgaag ggcgaacagt tcctgattaa ccacaaaccg ttctacttta
     1681 ctggctttgg tcgtcatgaa gatgcggact tgcgtggcaa aggattcgat aacgtgctga
     1741 tggtgcacga ccacgcatta atggactgga ttggggccaa ctcctaccgt acctcgcatt
     1801 acccttacgc tgaagagatg ctcgactggg cagatgaaca tggcatcgtg gtgattgatg
     1861 aaactgctgc tgtcggcttt aacctctctt taggcattgg tttcgaagcg ggcaacaagc
     1921 cgaaagaact gtacagcgaa gaggcagtca acggggaaac tcagcaagcg cacttacagg
     1981 cgattaaaga gctgatagcg cgtgacaaaa accacccaag cgtggtgatg tggagtattg
     2041 ccaacgaacc ggatacccgt ccgcaaggtg cacgggaata tttcgcgcca ctggcggaag
     2101 caacgcgtaa actcgacccg acgcgtccga tcacctgcgt caatgtaatg ttctgcgacg
     2161 ctcacaccga taccatcagc gatctctttg atgtgctgtg cctgaaccgt tattacggat
     2221 ggtatgtcca aagcggcgat ttggaaacgg cagagaaggt actggaaaaa gaacttctgg
     2281 cctggcagga gaaactgcat cagccgatta tcatcaccga atacggcgtg gatacgttag
     2341 ccgggctgca ctcaatgtac accgacatgt ggagtgaaga gtatcagtgt gcatggctgg
     2401 atatgtatca ccgcgtcttt gatcgcgtca gcgccgtcgt cggtgaacag gtatggaatt
     2461 tcgccgattt tgcgacctcg caaggcatat tgcgcgttgg cggtaacaag aaagggatct
     2521 tcactcgcga ccgcaaaccg aagtcggcgg cttttctgct gcaaaaacgc tggactggca
     2581 tgaacttcgg tgaaaaaccg cagcagggag gcaaacaacg cagggaggca aacaatgata
     2641 tcacaactct cctgacgcgt catcgtcggc tacagcctcg ggaattgcta cctagctcga
     2701 gcaagatcca aggagatata acaatggctt cctcctggat tgaacaagat ggattgcacg
     2761 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     2821 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca gggtagaccg gttctttttg
     2881 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     2941 ggctggccac gacgggcgta ccttgcgctg ctgtgctcga cgttgtcact gaagcgggaa
     3001 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3061 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3121 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3181 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3241 aactgttcgc caggctcaag gcgagaatgc ccgacggcga ggatctcgtc gtgacccatg
     3301 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3361 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3421 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3481 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga taaccgcgga
     3541 gagctcgaat ttccccgatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc
     3601 tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta agcatgtaat
     3661 aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca
     3721 attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc
     3781 gcgcgcggtg tcatctatgt tactagatcg tgggcccgtc cagttccatt cgccctatag
     3841 tgagtcgtat taaacctgca ttaatgaatc ggccaacgcg cggggtaggg ataacagggt
     3901 aatcccggga gagggtatga gaccactcat cgcgactagt agcggccgct gcagtccggc
     3961 aaaaaagggc aaggtgtcac caccctgccc tttttcttta aaaccgaaaa gattacttcg
     4021 cgttatgcag gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4081 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4141 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4201 ggcgtttttc cacaggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4261 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4321 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4381 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4441 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4501 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4561 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4621 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4681 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4741 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaagga
//
