LOCUS       sequence-397986-        3388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Even assembly vector with sfGFP dropout for Even level assemblies.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3388)
  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 3388)
  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_12 [18e4G]
FEATURES             Location/Qualifiers
     source          1..3388
                     /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"
     protein_bind    complement(2117..2141)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2241..3032,3033..3101))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2864..2883
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3102..3206)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3305..3327
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3319..3336
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3320..3336
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca gcttaatgtg aagagcgaaa 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 ctcttcaggt
     1081 acgcttgaga ccaccccagg 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 aaatcgtcct
     2101 cgtgatacca attcggagcc tgcttttttg tacaaacttg ttgataatgg caattcaagg
     2161 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2221 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2281 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2341 gagggcttac catctggccc cagtgctgca atgataccgc gagagccacg ctcaccggct
     2401 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2461 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2521 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     2581 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     2641 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     2701 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     2761 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     2821 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     2881 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     2941 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3001 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3061 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3121 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3181 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccaga tacctgaaac
     3241 aaaacccatc gtacggccaa ggaagtctcc aataactgtg atccaccaca agcgccaggg
     3301 ttttcccagt cacgacgttg taaaacgacg gccagtcatg cataatccgc acgcatctgg
     3361 aataaggaag tgccattccg cctgacct
//
