LOCUS       sequence-417378-        5879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 plasmid, barcoded ORI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5879)
  AUTHORS   Gilbert C, Lee H
  TITLE     POSSUM Expansion Pack
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5879)
  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            "unpublished"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: pGL1_107
FEATURES             Location/Qualifiers
     source          1..5879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(144..160)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(144..160)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(157..179)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(332..920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(412..431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1056..1871)
                     /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(1056..1871)
                     /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     1779..1798
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1863..1882
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1992..2008
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1992..2008
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      2009..2055
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2169..2191
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2183..2200
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2184..2200
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 agcatgaaga gcaccccagg atacatagat taccacaact ccgagccctt ccacctagcg
       61 tgctaaccac gcacgctatt gttgtatagg ctaggtggag gctcagtgat gataagtctg
      121 cgatggtgga tgcatgtgtc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
      181 agagggcaca atcctattcc gcgctatccg acaatctcca agacattagg tggagttcag
      241 ttcggcgtat ggcatatgtc gctggaaaga acatgtgagc aaaaggccag caaaaggcca
      301 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
      361 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
      421 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
      481 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
      541 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
      601 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
      661 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
      721 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
      781 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
      841 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
      901 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctctat
      961 tcaacaaagc cgccgtcccg tcaagtcagc gtaaatgggt agggggcttc aaatcgtccg
     1021 ctctgccagt gttacaacca attaacaaat tctgattaga aaaactcatc gagcatcaaa
     1081 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     1141 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     1201 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     1261 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc
     1321 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     1381 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     1441 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     1501 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     1561 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     1621 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     1681 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     1741 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     1801 tataaatcag catccatgtt ggaatttaat cgcggcctcg agcaagacgt ttcccgttga
     1861 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     1921 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt
     1981 tcccccgccg ctctagaact agtggatcca aataaaacga aaggctcagt cgaaagactg
     2041 ggcctttcgt tttatctgtt gtttgtcgca ttatacgaga cgtccaggtt gggatacctg
     2101 aaacaaaacc catcgtacgg ccaaggaagt ctccaataac tgtgatccac cacaagcgcc
     2161 agggttttcc cagtcacgac gttgtaaaac gacggccagt catgcataat ccgcacgcat
     2221 ctggaataag gaagtgccat tccgcctgac cttagcgtgc gaacagcacg ctattgttgt
     2281 atcattactc gcatccattc tcaggctgtc tcgtctcgtc tccgagctct tcaatgagtc
     2341 agggtcacgc gtaggacgcg tcgagtaggg ttaaatcgtg gatggaggag ggttccgcag
     2401 ccacacgccg tgtgaagctg gggagggcgc gcccgccctt aagtctaaaa attaggggag
     2461 atgtaaggat ttgggaaaaa tagaagatgt tataatcata aatatggtat tcgtaggctt
     2521 aaagtcaaaa aggaggtgaa atataaatag atttttagct aaattaagta agaaatagga
     2581 ggagatttat tgaacaaaaa attagaaaaa ccatttgtat ataagagaga gtacgatttg
     2641 actggatatg atgttgaaat tttacaaaaa tatgagttag aacaagcaat atatgtttat
     2701 gttgggagta gttgtgcata taacatgaga gctagaagta gtaaatggag ataccatata
     2761 agaacaaata ataagtctat atgttgtaac attaaaaatt ttatacataa cttggaattg
     2821 ttttataaaa tggaattaaa gttgtcagat aatattatta atgataagct atactatagc
     2881 aatatagcag agtttgaaga atttgaaaca ctagaaaaag ctagagaggt agaaagtact
     2941 attataagtc aatatcaatt tttagattct ataaatcaca tgttaaaaca aaaaataatt
     3001 ttattgagta ataaggatag tgtgttaaac ataactaaaa atggaaatac aaattatttg
     3061 aaagtaaaaa ataaatacat agaaaaacat aagaacaagc caataatgag ataccatatc
     3121 aactgtcaat tcaatacaga tggaagtgtc aaaagtatta cacaggagtt tgaaccaata
     3181 ttggaattaa acaaaaaaaa taccctaagc cgaccaagca gagtattttt aaaataatat
     3241 tttaagataa caacaaaatg agataatact actagacaat gacaactcaa ctaccaattg
     3301 agtttatgga gctaccaact ccaatatcgg tctaactgat taagtatctg tagttatata
     3361 ataatattgc tatcaatttt agcatcttaa caatattatt atacatacta agctaaaatt
     3421 attcaatagt tgtaaaagtt gattagtcaa taagtatata tttaatgtag tgttatctct
     3481 taaaaaaact agataaggag ataataaata tatggaacaa ttagattcaa aatataagtt
     3541 gaaaaaattt ctaatggcag tatttagaga tggtatagga caaggaaata atcttattga
     3601 taatgaatat gttagagtat ttcaaaataa taaaagtaat agtaaacaat tagaactcgg
     3661 agaagaattt aaagaatata gtaaaacaac tttttttaaa aatatagatg atatagtaga
     3721 atttaccttc gcaaaaaata tttattatga aaatacattt tttaacctat gtactactga
     3781 tggaaaagca ggaaccaatg aaaacttaat aaatagatat gcattaggat ttgattttga
     3841 caaaaaagaa ttaggacaag gttttaatta taaagatata attaatttat ttactaagat
     3901 aggattacat tatcatatcc tagttgatag tggaaatgga ttccatgttt atgtgctaat
     3961 taataaaact aataacatta agttagtatc agaagttaca aatacattaa taaataaatt
     4021 gggtgcagat aaacaagcaa atttatctac tcaagtatta agagtacctt atacatataa
     4081 tattaaaaat actactaaac aagtaaaaat aatacaccaa gacaaaaata tatatagata
     4141 tgacatagaa aagttagcta aaaaatattg caaagatgta aaaacagtag gtaatactaa
     4201 tacaaaatat atattagata gtaagctacc aaattgtata gtagatattt taaaaaatgg
     4261 tagtaaagat ggacataaaa acctagattt gcaaaaaata gttgtgactt taagattgag
     4321 gaataaaagt ttaagtcaag taatatccgt tgctagagaa tggaactata tatcacaaaa
     4381 tagtctttca aatagtgagc tagaatatca agtcaagtat atgtatgaga aacttaaaac
     4441 ggttaatttt ggttgtactg gttgtgagtt taatagtgat tgttggaata aaatagaatc
     4501 agattttata tatagtgatg aagatacttt gttcaatatg ccacataagc actcaaagga
     4561 tttgaaatat aagaatagga aaggggttaa aataatgact ggtaatcaat tgtttatcta
     4621 taatgtgtta cttaacaata aagatagaga attaaacata gacgatataa tggagctgat
     4681 aacctataaa cgtaagaaga aagttaaaaa cattgttatg agtgaaaaga cattaagaga
     4741 aacattaaaa gaacttcaac ataatgatta tattacaaaa acaaaaggtg ttacaaagct
     4801 aggaataaaa gatacataca atgtaaaaga agttagatgt aatatagata aacaatatac
     4861 tattagttac tttgttacca tggcagtaat ttggggaata atttcaactg aagaattaag
     4921 attatatact cacatgagat ataagcaaga tttattggtc aaagatgata aaataaaagg
     4981 aaatatatta agaattaatc aagaggaatt agcaaaagat ttaggagtaa cacagcaaag
     5041 aatttcaaat atgatagaat ctttattaga tactaaaatt ttagatgtat gggaaactaa
     5101 aataaatgat agaggattta tgtactatac atatagatta aacaagtaga tttttgatag
     5161 gattagaatt gattttctag tcctattttt atgcaaaaaa actaattaat aaaaatttct
     5221 tttggtaaaa taattgtacg agaattgcaa aaaaaaaatg gcatcaaagt attgaaatta
     5281 agccgtttta aaaatttctt ttggtaaaat aattctacat atatatgtag tatatatata
     5341 tatgtttttt agagaatgta taactagaat atagagctag aatatagaga atgtataact
     5401 agaatataga gctagaatat agagaatgta taactagaat atagagctag aatatagaga
     5461 atgtataact agaatataga gctagaatat agagaatgta taactagaat atagagctag
     5521 aatatagaga atgtataact agaatataga gctagaatat agagaatgta taactagaat
     5581 atagagctag aatcctaatt agtaggtgct tttttaaaac aagttaaaaa tcaaaaatag
     5641 tattagtaag cattggaaat gctagattct aaaatagaaa agtaaaaaat tggtgcacta
     5701 tctaaactta tctatatcgc tttttccgtc gtttggttct ctagttacga tacaggggat
     5761 atgcttatat tgagttatag tactaatcag tgcttaatat agttaataaa attatagtta
     5821 ccatagttta gtaactatga tgtatgttag ttagaaactt gcatttcggc cggcccgct
//
