LOCUS       sequence-252100-        3870 bp    DNA     circular SYN 15-DEC-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 3870)
  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: "Synth Biol 
            (Oxf)."; date: "2021-02-2"; volume: "6(1):ysab003. doi"; pages: " 
            10.1093/synbio/ysab003. eCollection 2021"
COMMENT     Sequence Label: pJUMP49-2B(sfGFP)
FEATURES             Location/Qualifiers
     source          1..3870
                     /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    926..946
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      980..1074
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1310..2101
                     /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            2247..2355
                     /note="incP origin of transfer"
     rep_origin      2391..2979
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2880..2899
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3133..3150
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3165..3305
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3231..3250)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3391..3413
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3407..3598)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      complement(3778..3864)
                     /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 ggtctctaat 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 cgctagcctg agacctacta gtagcggccg ctgcagggag ttgtcttcga
      961 agacttcgct ctagtcttgg actcctgttg atagatccag taatgacctc agaactccat
     1021 ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaggg
     1081 gtccccaata attacgattt acgtatttaa atgaaccttg accgaacgca gcggtggtaa
     1141 cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc
     1201 tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag
     1261 caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaaacatca tgagggaagc
     1321 ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct
     1381 cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc
     1441 acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg
     1501 agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg
     1561 cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa
     1621 gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc
     1681 agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc
     1741 cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga
     1801 ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gtgatgagcg
     1861 aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc
     1921 gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat
     1981 acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga
     2041 tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata
     2101 agacaattgt ccttttccgc tgcataaccc tgcttcgggg tcattatagc gattttttcg
     2161 gtatatccat cctttttcgc acgatataca ggattttgcc aaagggttcg tgtagacttt
     2221 ccttggtgta tccaacggcg tcagccgggc aggataggtg aagtaggccc acccgcgagc
     2281 gggtgttcct tcttcactgt cccttattcg cacctggcgg tgctcaacgg gaatcctgct
     2341 ctgcgaggct ggccgtaggc cggccccgta gaaaagatca aaggatcttc ttgagatcct
     2401 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2461 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2521 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     2581 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2641 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2701 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2761 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2821 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2881 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2941 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3001 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3061 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3121 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     3181 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc
     3241 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     3301 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3361 catccgctta cagacaagct gtgacagtct ccgggagctg catgtgtcag aggttttcac
     3421 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgcagcg
     3481 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     3541 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     3601 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     3661 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     3721 gtaaacaact ggcggtatgg atgcggcggg ggcgcgccca gctgtctagg gcggcggatt
     3781 tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttt
     3841 cgactgagcc tttcgtttta tttgatgcct
//
