LOCUS       sequence-255059-        3866 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Level 2 vector. OriV 9 (pBBR322/ROP; medium copy number); sfGFP 
            cloning reporter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 87 to 121)
  AUTHORS   Valenzuela-Ortega M, French C
  TITLE     Joint universal modular plasmids (JUMP): a flexible vector platform 
            for synthetic biology.
  JOURNAL   Synth Biol (Oxf). 2021 Feb 2;6(1):ysab003. doi: 
            10.1093/synbio/ysab003. eCollection 2021.
  PUBMED    33623824
REFERENCE   2  (bases 1 to 3866)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Synth Biol 
            (Oxf)."; date: "2021-02-2"; volume: "6(1):ysab003. doi"; pages: " 
            10.1093/synbio/ysab003. eCollection 2021"
COMMENT     Sequence Label: pJUMP49-2D(sfGFP)
FEATURES             Location/Qualifiers
     source          1..3866
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    39..60
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        87..121
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     RBS             126..146
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             163..873
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     misc_feature    922..942
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      976..1070
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1306..2097
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     oriT            2243..2351
                     /note="incP origin of transfer"
     rep_origin      2387..2975
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2876..2895
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3129..3146
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3161..3301
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3227..3246)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3387..3409
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3403..3594)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      complement(3774..3860)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaattaagg agttttgcag gtgcacctgc ttttcgctga attcgcggcc gcttctagag
       61 ggtctctttc gggagtgaga cgggagttga cggctagctc agtcctaggt acagtgctag
      121 cgtacttgtt taactttaag aaggagatat acaatggtag cccgtaaagg cgaagagctg
      181 ttcactggtg tcgtccctat tctggtggaa ctggatggtg atgtcaacgg tcataagttt
      241 tccgtgcgtg gcgagggtga aggtgacgca actaatggta aactgacgct gaagttcatc
      301 tgtactactg gtaaactgcc ggtaccttgg ccgactctgg taacgacgct gacttatggt
      361 gttcagtgct ttgctcgtta tccggaccat atgaagcagc atgacttctt caagtccgcc
      421 atgccggaag gctatgtgca ggaacgcacg atttccttta aggatgacgg cacgtacaaa
      481 acgcgtgcgg aagtgaaatt tgaaggcgat accctggtaa accgcattga gctgaaaggc
      541 attgacttta aagaagatgg caatatcctg ggccataagc tggaatacaa ttttaacagc
      601 cacaatgttt acatcaccgc cgataaacaa aaaaatggca ttaaagcgaa ttttaaaatt
      661 cgccacaacg tggaggatgg cagcgtgcag ctggctgatc actaccagca aaacactcca
      721 atcggtgatg gtcctgttct gctgccagac aatcactatc tgagcacgca aagcgttctg
      781 tctaaagatc cgaacgagaa acgcgatcat atggttctgc tggagttcgt aaccgcagcg
      841 ggcatcacgc atggtatgga tgaactgtac aaaggttcgt aacttctgac tgagttgcac
      901 gctcgtctct cgcttgagac ctactagtag cggccgctgc agggagttgt cttcgaagac
      961 ttcgctctag tcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg
     1021 atttgttcag aacgctcggt tgccgccggg cgttttttat tggtgagaat ccaggggtcc
     1081 ccaataatta cgatttacgt atttaaatga accttgaccg aacgcagcgg tggtaacggc
     1141 gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc tatgcctcgg
     1201 gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac
     1261 gatgttacgc agcagggcag tcgccctaaa acaaagttaa acatcatgag ggaagcggtg
     1321 atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa
     1381 ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac
     1441 agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct
     1501 ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct
     1561 gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc
     1621 gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc
     1681 acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg
     1741 gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg
     1801 ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggtga tgagcgaaat
     1861 gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag
     1921 gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt
     1981 gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag
     2041 ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataagac
     2101 aattgtcctt ttccgctgca taaccctgct tcggggtcat tatagcgatt ttttcggtat
     2161 atccatcctt tttcgcacga tatacaggat tttgccaaag ggttcgtgta gactttcctt
     2221 ggtgtatcca acggcgtcag ccgggcagga taggtgaagt aggcccaccc gcgagcgggt
     2281 gttccttctt cactgtccct tattcgcacc tggcggtgct caacgggaat cctgctctgc
     2341 gaggctggcc gtaggccggc cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2401 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2461 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2521 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2581 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2641 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2701 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2761 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2821 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2881 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2941 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3001 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3061 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     3121 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     3181 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     3241 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     3301 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3361 cgcttacaga caagctgtga cagtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3421 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gcagcgattc
     3481 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     3541 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     3601 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     3661 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     3721 acaactggcg gtatggatgc ggcgggggcg cgcccagctg tctagggcgg cggatttgtc
     3781 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtctttcgac
     3841 tgagcctttc gttttatttg atgcct
//
