LOCUS       sequence-417391-        5130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Even ORI-marker screen destination vector; contains sfYFP dropout 
            for barcoded-ORI insertion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 3650 to 3684)
  AUTHORS   Gilbert C, Lee H
  TITLE     POSSUM Expansion Pack
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5130)
  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: 
            "unpublished"
COMMENT     Sequence Label: pGL2_212
FEATURES             Location/Qualifiers
     source          1..5130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(469..493)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(593..1384,1385..1453))
                     /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     1216..1235
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1454..1558)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1657..1679
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1671..1688
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1672..1688
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             1988..1999
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2006..2719
                     /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
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     promoter        3650..3684
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     oriT            4610..4718
                     /note="incP origin of transfer"
     primer_bind     complement(4872..4888)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4872..4888)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4885..4907)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      join(5078..5130,1..336)
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 gggcttctca gtgcgttaca tccctggctt gttgtccaca accgttaaac cttaaaggct
       61 ttaaaagcct tatatattct tttttttctt ataaaactta aaaccttaga ggctatttaa
      121 gttgctgatt tatattaatt ttattgttca aacatgagag cttagtacgt gaaacatgag
      181 agcttagtac gttagccatg agagcttagt acgttagcca tgagggttta gttcgttaaa
      241 catgagagct tagtacgtta aacatgagag cttagtacgt gaaacatgag agcttagtac
      301 gtactatcaa caggttgaac tgctgatctt cagatcggcg cgccggccgg ccgaagatcc
      361 tttgatcttt tctacggggt ctgacgctct attcaacaaa gccgccgtcc cgtcaagtca
      421 gcgtaaatgg gtagggggct tcaaatcgtc ctcgtgatac caattcggag cctgcttttt
      481 tgtacaaact tgttgataat ggcaattcaa ggatcttcac ctagatcctt ttaaattaaa
      541 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
      601 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
      661 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
      721 caatgatacc gcgagagcca cgctcaccgg ctccagattt atcagcaata aaccagccag
      781 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
      841 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
      901 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
      961 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     1021 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     1081 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1141 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     1201 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     1261 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     1321 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     1381 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     1441 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     1501 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     1561 catttccccg aaaagtgcca gatacctgaa acaaaaccca tcgtacggcc aaggaagtct
     1621 ccaataactg tgatccacca caagcgccag ggttttccca gtcacgacgt tgtaaaacga
     1681 cggccagtca tgcataatcc gcacgcatct ggaataagga agtgccattc cgcctgacct
     1741 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
     1801 gtctcgtctc cgaggtctca ggagaatgtg aagagcttcg aaagtgaaac gtgatttcat
     1861 gcgtcatttt gaacattttg taaatcttat ttaataatgt gtgcggcaat tcacatttaa
     1921 tttatgaatg ttttcttaac atcgcggcaa ctcaagaaac ggcaggttcg gatcttagct
     1981 actagagaaa gaggagaaat actagatgcg taaaggcgaa gaactgttca ctggtgtcgt
     2041 ccctattctg gtggaactgg atggtgatgt caacggtcat aagttttccg tgcgtggcga
     2101 gggtgaaggt gacgcaacta atggtaaact gacgctgaag ttcatctgta ctactggtaa
     2161 actgccggta ccttggccga ctctggtaac gacgctgact tatggtgttc agtgctttgc
     2221 tcgttatccg gaccatatga agcagcatga cttcttcaag tccgccatgc cggaaggcta
     2281 tgtgcaggaa cgcacgattt cctttaagga tgacggcacg tacaaaacgc gtgcggaagt
     2341 gaaatttgaa ggcgataccc tggtaaaccg cattgagctg aaaggcattg actttaaaga
     2401 agacggcaat atcctgggcc ataagctgga atacaatttt aacagccaca atgtttacat
     2461 caccgccgat aaacaaaaaa atggcattaa agcgaatttt aaaattcgcc acaacgtgga
     2521 ggatggcagc gtgcagctgg ctgatcacta ccagcaaaac actccaatcg gtgatggtcc
     2581 tgttctgctg ccagacaatc actatctgag ctaccaaagc gttctgtcta aagatccgaa
     2641 cgagaaacgc gatcatatgg ttctgctgga gttcgtaacc gcagcgggca tcacgcatgg
     2701 tatggatgaa ctgtacaaat aattcagcca aaaaacttaa gaccgccggt cttgtccact
     2761 accttgcagt aatgcggtgg acaggatcgg cggttttctt ttctcttctc aagaagctct
     2821 tcagcaagga gggccggcca gtgggcaagt tgaaaaattc acaaaaatgt ggtataatat
     2881 ctttgttcat tagagcgata aacttgaatt tgagagggaa cttagaatgg tatttgaaaa
     2941 aattgataaa aatagttgga acagaaaaga gtattttgac cactactttg caagtgtacc
     3001 ttgtacctac agcatgaccg ttaaagtgga tatcacacaa ataaaggaaa agggaatgaa
     3061 actatatcct gcaatgcttt attatattgc aatgattgta aaccgccatt cagagtttag
     3121 gacggcaatc aatcaagatg gtgaattggg gatatatgat gagatgatac caagctatac
     3181 aatatttcac aatgatactg aaacattttc cagcctttgg actgagtgta agtctgactt
     3241 taaatcattt ttagcagatt atgaaagtga tacgcaacgg tatggaaaca atcatagaat
     3301 ggaaggaaag ccaaatgctc cggaaaacat ttttaatgta tctatgatac cgtggtcaac
     3361 cttcgatggc tttaatctga atttgcagaa aggatatgat tatttgattc ctatttttac
     3421 tatggggaaa tattataaag aagataacaa aattatactt cctttggcaa ttcaagttca
     3481 tcacgcagta tgtgacggat ttcacatttg ccgttttgta aacgaattgc aggaattgat
     3541 aaatagttaa gcttcaggtt tgtctgtaac taaaaacaag tatttaagca aaaacatcgt
     3601 agaaatacgg tgttttttgt taccctaagt ttaaaccgct atacaggagt tgacggctag
     3661 ctcagtccta ggtacagtgc tagctactag gaggaaaaaa atggtgagta agggtgaagc
     3721 cgtgattaaa gagttcatgc gcttcaaagt gcatatggag ggctcaatga acggccatga
     3781 atttgaaatt gaaggtgaag gagaaggccg tccgtatgag ggcacccaga ccgccaaact
     3841 gaaagtcaca aagggtggcc cgctgccgtt ttcgtgggat attttatccc ctcaattcat
     3901 gtacggcagc cgggcgttta ttaaacaccc ggcggatatc ccggattact ataagcaatc
     3961 ttttccagaa ggctttaaat gggagcgcgt gatgaatttt gaagacggag gtgctgttac
     4021 cgtgacccag gacacgagct tagaagacgg taccttgatc tataaggtaa agctgcgtgg
     4081 taccaatttc ccgccggatg ggccagttat gcagaaaaag actatgggct gggaagcatc
     4141 gacggagcgt ctttaccccg aagacggcgt cctgaaaggc gatattaaaa tggccctgcg
     4201 actgaaagac ggcggtcgct accttgcaga tttcaaaaca acgtataaag cgaaaaaacc
     4261 tgtgcagatg cccggggcgt acaacgttga tcgcaaacta gatatcactt cacacaacga
     4321 agattacacg gtagtcgaac agtatgaacg tagcgagggg cgccatagca ccggaggcat
     4381 ggatgaactc tataaataag cttcgctcgt gctcgccgcg accttcctct tcatgcacct
     4441 cttcggcatc gggctgcaca acgctacaga ggagcttttc cgctgcataa ccctgcttcg
     4501 gggtcattat agcgattttt tcggtatatc catccttttt cgcacgatat acaggatttt
     4561 gccaaagggt tcgtgtagac tttccttggt gtatccaacg gcgtcagccg ggcaggatag
     4621 gtgaagtagg cccacccgcg agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg
     4681 cggtgctcaa cgggaatcct gctctgcgag gctggccgta cgctaggtat acttgagacc
     4741 accccaggat acatagatta ccacaactcc gagcccttcc acctagcgtg ctaaccacgc
     4801 acgctattgt tgtataggct aggtggaggc tcagtgatga taagtctgcg atggtggatg
     4861 catgtgtcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcag agggcacaat
     4921 cctattccgc gctatccgac aatctccaag acattaggtg gagttcagtt cggcgtatgg
     4981 catatgtcgc tggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5041 aggccgcgtt gctggcgtta cagctgggcg cgccccatgt cagccgttaa gtgttcctgt
     5101 gtcactcaaa attgctttga gaggctctaa
//
