LOCUS       sequence-397999-        3332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Odd assembly vector with sfGFP dropout for Odd level assemblies.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3332)
  AUTHORS   Gilbert C, Brumwell SL, Crits-Christoph A, Kang SC, Martin-Moldes Z,
            Alsharif W, Esmurria A, Nguyen M-A, Lee HH, Ostrov N
  TITLE     A scalable framework for high-throughput identification of 
            functional origins of replication in non-model bacteria
  JOURNAL   bioRxiv 2023.05.19.541510
REFERENCE   2  (bases 1 to 3332)
  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: "bioRxiv 
            2023.05.19.541510"
COMMENT     Sequence Label: pGLDV_8 [28o4G]
FEATURES             Location/Qualifiers
     source          1..3332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             245..256
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             263..976
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1224..1240)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1224..1240)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1237..1259)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1412..2000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1492..1511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2136..2951)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2136..2951)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2859..2878
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2943..2962
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3072..3088
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3072..3088
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      3089..3135
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3249..3271
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3263..3280
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3264..3280
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgagctcttc acagaggagt gagaccgaaa gtgaaacgtg atttcatgcg
      121 tcattttgaa cattttgtaa atcttattta ataatgtgtg cggcaattca catttaattt
      181 atgaatgttt tcttaacatc gcggcaactc aagaaacggc aggttcggat cttagctact
      241 agagaaagag gagaaatact agatgcgtaa aggcgaagaa ctgttcactg gtgtcgtccc
      301 tattctggtg gaactggatg gtgatgtcaa cggtcataag ttttccgtgc gtggcgaggg
      361 tgaaggtgac gcaactaatg gtaaactgac gctgaagttc atctgtacta ctggtaaact
      421 gccggttcct tggccgactc tggtaacgac gctgacttat ggtgttcagt gctttgctcg
      481 ttatccggac catatgaagc agcatgactt cttcaagtcc gccatgccgg aaggctatgt
      541 gcaggaacgc acgatttcct ttaaggatga cggcacgtac aaaacgcgtg cggaagtgaa
      601 atttgaaggc gataccctgg taaaccgcat tgagctgaaa ggcattgact ttaaagagga
      661 cggcaatatc ctgggccata agctggaata caattttaac agccacaatg tttacatcac
      721 cgccgataaa caaaaaaatg gcattaaagc gaattttaaa attcgccaca acgtggagga
      781 tggcagcgtg cagctggctg atcactacca gcaaaacact ccaatcggtg atggtcctgt
      841 tctgctgcca gacaatcact atctgagcac gcaaagcgtt ctgtctaaag atccgaacga
      901 gaaacgcgat catatggttc tgctggagtt cgtaaccgca gcgggcatca cgcatggtat
      961 ggatgaactg tacaaatgat tcagccaaaa aacttaagac cgccggtctt gtccactacc
     1021 ttgcagtaat gcggtggaca ggatcggcgg ttttcttttc tcttctcaag gtctcacgct
     1081 aggttgaaga gcaccccagg atacatagat taccacaact ccgagccctt ccacctagcg
     1141 tgctaaccac gcacgctatt gttgtatagg ctaggtggag gctcagtgat gataagtctg
     1201 cgatggtgga tgcatgtgtc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1261 agagggcaca atcctattcc gcgctatccg acaatctcca agacattagg tggagttcag
     1321 ttcggcgtat ggcatatgtc gctggaaaga acatgtgagc aaaaggccag caaaaggcca
     1381 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1441 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1501 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1561 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1621 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1681 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1741 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     1801 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     1861 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1921 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1981 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctctat
     2041 tcaacaaagc cgccgtcccg tcaagtcagc gtaaatgggt agggggcttc aaatcgtccg
     2101 ctctgccagt gttacaacca attaacaaat tctgattaga aaaactcatc gagcatcaaa
     2161 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     2221 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     2281 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     2341 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc
     2401 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     2461 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     2521 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     2581 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     2641 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     2701 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     2761 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     2821 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     2881 tataaatcag catccatgtt ggaatttaat cgcggcctcg agcaagacgt ttcccgttga
     2941 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     3001 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt
     3061 tcccccgccg ctctagaact agtggatcca aataaaacga aaggctcagt cgaaagactg
     3121 ggcctttcgt tttatctgtt gtttgtcgca ttatacgaga cgtccaggtt gggatacctg
     3181 aaacaaaacc catcgtacgg ccaaggaagt ctccaataac tgtgatccac cacaagcgcc
     3241 agggttttcc cagtcacgac gttgtaaaac gacggccagt catgcataat ccgcacgcat
     3301 ctggaataag gaagtgccat tccgcctgac ct
//
