LOCUS       sequence-398503-        3222 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 3222)
  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 3222)
  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_19 [11e1G]
FEATURES             Location/Qualifiers
     source          1..3222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     305..324
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(543..647)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     746..768
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     760..777
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     761..777
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             1074..1085
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1092..1805
                     /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(2053..2069)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2053..2069)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2066..2088)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      2259..2647
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(2780..2804)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2904..3222,1..473,474..542))
                     /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"
ORIGIN
        1 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
       61 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
      121 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
      181 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
      241 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
      301 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
      361 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
      421 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
      481 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
      541 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
      601 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
      661 aaagtgccag atacctgaaa caaaacccat cgtacggcca aggaagtctc caataactgt
      721 gatccaccac aagcgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtcat
      781 gcataatccg cacgcatctg gaataaggaa gtgccattcc gcctgacctt agcgtgcgaa
      841 cagcacgcta ttgttgtatc attactcgca tccattctca ggctgtctcg tctcgtctcc
      901 gaggtctcag gagaatgtga agagcgaaag tgaaacgtga tttcatgcgt cattttgaac
      961 attttgtaaa tcttatttaa taatgtgtgc ggcaattcac atttaattta tgaatgtttt
     1021 cttaacatcg cggcaactca agaaacggca ggttcggatc ttagctacta gagaaagagg
     1081 agaaatacta gatgcgtaaa ggcgaagaac tgttcactgg tgtcgtccct attctggtgg
     1141 aactggatgg tgatgtcaac ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg
     1201 caactaatgg taaactgacg ctgaagttca tctgtactac tggtaaactg ccggttcctt
     1261 ggccgactct ggtaacgacg ctgacttatg gtgttcagtg ctttgctcgt tatccggacc
     1321 atatgaagca gcatgacttc ttcaagtccg ccatgccgga aggctatgtg caggaacgca
     1381 cgatttcctt taaggatgac ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg
     1441 ataccctggt aaaccgcatt gagctgaaag gcattgactt taaagaggac ggcaatatcc
     1501 tgggccataa gctggaatac aattttaaca gccacaatgt ttacatcacc gccgataaac
     1561 aaaaaaatgg cattaaagcg aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc
     1621 agctggctga tcactaccag caaaacactc caatcggtga tggtcctgtt ctgctgccag
     1681 acaatcacta tctgagcacg caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc
     1741 atatggttct gctggagttc gtaaccgcag cgggcatcac gcatggtatg gatgaactgt
     1801 acaaatgatt cagccaaaaa acttaagacc gccggtcttg tccactacct tgcagtaatg
     1861 cggtggacag gatcggcggt tttcttttct cttctcaagc tcttcaggta tacttgagac
     1921 caccccagga tacatagatt accacaactc cgagcccttc cacctagcgt gctaaccacg
     1981 cacgctattg ttgtataggc taggtggagg ctcagtgatg ataagtctgc gatggtggat
     2041 gcatgtgtca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca gagggcacaa
     2101 tcctattccg cgctatccga caatctccaa gacattaggt ggagttcagt tcggcgtatg
     2161 gcatatgtcg ctggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2221 aaggccgcgt tgctggcgtt acagctgggc gcgccccatg tcagccgtta agtgttcctg
     2281 tgtcactcaa aattgctttg agaggctcta agggcttctc agtgcgttac atccctggct
     2341 tgttgtccac aaccgttaaa ccttaaaggc tttaaaagcc ttatatattc ttttttttct
     2401 tataaaactt aaaaccttag aggctattta agttgctgat ttatattaat tttattgttc
     2461 aaacatgaga gcttagtacg tgaaacatga gagcttagta cgttagccat gagagcttag
     2521 tacgttagcc atgagggttt agttcgttaa acatgagagc ttagtacgtt aaacatgaga
     2581 gcttagtacg tgaaacatga gagcttagta cgtactatca acaggttgaa ctgctgatct
     2641 tcagatcggc gcgccggccg gccgaagatc ctttgatctt ttctacgggg tctgacgctc
     2701 tattcaacaa agccgccgtc ccgtcaagtc agcgtaaatg ggtagggggc ttcaaatcgt
     2761 cctcgtgata ccaattcgga gcctgctttt ttgtacaaac ttgttgataa tggcaattca
     2821 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2881 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2941 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3001 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagagcc acgctcaccg
     3061 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3121 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3181 tcgccagtta atagtttgcg caacgttgtt gccattgcta ca
//
