LOCUS       sequence-417373-        4836 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 4836)
  AUTHORS   Gilbert C, Lee H
  TITLE     POSSUM Expansion Pack
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 4836)
  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            "unpublished"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_105
FEATURES             Location/Qualifiers
     source          1..4836
                     /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 tttataatca tcctccccgg cgttccgcag
     2401 ccacacgccg tgtgaagctg gggagggcgc gccgttctga atccttagct aatggttcaa
     2461 caggtaacta tgacgaagat agcaccctgg ataagtctgt aatggattct aaggcattta
     2521 atgaagacgt gtatataaaa tgtgctaatg aaaaagaaaa tgcgttaaaa gagcctaaaa
     2581 tgagttcaaa tggttttgaa attgattggt agtttaattt aatatatttt ttctattggc
     2641 tatctcgata cctatagaat cttctgttca cttttgtttt tgaaatataa aaaggggctt
     2701 tttagcccct tttttttaaa actccggagg agtttcttca ttcttgatac tatacgtaac
     2761 tattttcgat ttgacttcat tgtcaattaa gctagtaaaa tcaatggtta aaaaacaaaa
     2821 aacttgcatt tttctaccta gtaatttata attttaagtg tcgagtttaa aagtataatt
     2881 taccaggaaa ggagcaagtt ttttaataag gaaaaatttt tccttttaaa attctatttc
     2941 gttatatgac taattataat caaaaaaatg aaaataaaca agaggtaaaa actgctttag
     3001 agaaatgtac tgataaaaaa agaaaaaatc ctagatttac gtcatacata gcacctttaa
     3061 ctactaagaa aaatattgaa aggacttcca cttgtggaga ttatttgttt atgttgagtg
     3121 atgcagactt agaacatttt aaattacata aaggtaattt ttgcggtaat agattttgtc
     3181 caatgtgtag ttggcgactt gcttgtaagg atagtttaga aatatctatt cttatggagc
     3241 atttaagaaa agaagaaaat aaagagttta tatttttaac tcttacaact ccaaatgtaa
     3301 aaagttatga tcttaattat tctattaaac aatataataa atcttttaaa aaattaatgg
     3361 agcgtaagga agttaaggat ataactaaag gttatataag aaaattagaa gtaacttacc
     3421 aaaaggaaaa atacataaca aaggatttat ggaaaataaa aaaagattat tatcaaaaaa
     3481 aaggacttga aattggtgat ttagaaccta attttgatac ttataatcct cattttcatg
     3541 tagttattgc agttaataaa agttatttta cagataaaaa ttattatata aatcgagaaa
     3601 gatggttgga attatggaag tttgctacta aggatgattc tataactcaa gttgatgtta
     3661 gaaaagcaaa aattaatgat tataaagagg tttacgaact tgcgaaatat tcagctaaag
     3721 acactgatta tttaatatcg aggccagtat ttgaaatttt ttataaagca ttaaaaggca
     3781 agcaggtatt agtttttagt ggatttttta aagatgcaca caaattgtac aagcaaggaa
     3841 aacttgatgt ttataaaaag aaagatgaaa ttaaatatgt ctatatagtt tattataatt
     3901 ggtgcaaaaa acaatatgaa aaaactagaa taagggaact tacggaagat gaaaaagaag
     3961 aattaaatca agatttaata gatgaaatag aaatagatta aagtgtaact atactttata
     4021 tatatatgat taaaaaaata aaaaacaaca gcctattagg ttgttgtttt ttattttctt
     4081 tattaatttt tttaattttt agtttttagt tcttttttaa aataagtttc agcctctttt
     4141 tcaatatttt ttaaagaagg agtatttgca tgaattgcct tttttctaac agacttagga
     4201 aatattttaa cagtatcttc ttgcgccggt gattttggaa cttcataact tactaattta
     4261 taattattat tttctttttt aattgtaaca gttgcaaaag aagctgaacc tgttccttca
     4321 actagtttat catcttcaat ataatattct tgacctatat agtataaata tatttttatt
     4381 atatttttac ttttttctga atctattatt ttataatcat aaaaagtttt accaccaaaa
     4441 gaaggttgta ctccttctgg tccaacatat ttttttacta tattatctaa ataatttttg
     4501 ggaactggtg ttgtaatttg attaatcgaa caaccagtta tacttaaagg aattataact
     4561 ataaaaatat ataggattat ctttttaaat ttcattattg gcctcctttt tattaaattt
     4621 atgttaccat aaaaaggaca taacgggaat atgtagaata tttttaatgt agacaaaatt
     4681 ttacataaat ataaagaaag gaagtgtttg tttaaatttt atagcaaact atcaaaaatt
     4741 agggggataa aaatttatga aaaaaaggtt ttcgatgtta tttttatgtt taactttaat
     4801 agtttgtggt ttatttacaa attcggccgg cccgct
//
