LOCUS       sequence-417295-        5248 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 part.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5248)
  AUTHORS   Gilbert C, Lee H
  TITLE     POSSUM Expansion Pack
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5248)
  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            "unpublished"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: pGL0_152 [pBP1 ORI]
FEATURES             Location/Qualifiers
     source          1..5248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3027..3043)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3027..3043)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3040..3062)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3051..3067
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3075..3081,3082..3099,3100..3105))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3120..3141
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3258..3275)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3429..4017)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3509..3528)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4188..4979,4980..5048))
                     /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     4811..4830
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5049..5153)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5221..5239
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagactgtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gacgcgtatt gggattagcg
      421 tgcgaacagc acgctattgt tgtatcatta ctcgcatcca ttctcaggct gtctcgtctc
      481 gtctccgagg tctcaggagg gcgcgccgtt ctgaatcctt agctaatggt tcaacaggta
      541 actatgacga agatagcacc ctggataagt ctgtaatgga ttctaaggca tttaatgaag
      601 acgtgtatat aaaatgtgct aatgaaaaag aaaatgcgtt aaaagagcct aaaatgagtt
      661 caaatggttt tgaaattgat tggtagttta atttaatata ttttttctat tggctatctc
      721 gatacctata gaatcttctg ttcacttttg tttttgaaat ataaaaaggg gctttttagc
      781 cccttttttt taaaactccg gaggagtttc ttcattcttg atactatacg taactatttt
      841 cgatttgact tcattgtcaa ttaagctagt aaaatcaatg gttaaaaaac aaaaaacttg
      901 catttttcta cctagtaatt tataatttta agtgtcgagt ttaaaagtat aatttaccag
      961 gaaaggagca agttttttaa taaggaaaaa tttttccttt taaaattcta tttcgttata
     1021 tgactaatta taatcaaaaa aatgaaaata aacaagaggt aaaaactgct ttagagaaat
     1081 gtactgataa aaaaagaaaa aatcctagat ttacgtcata catagcacct ttaactacta
     1141 agaaaaatat tgaaaggact tccacttgtg gagattattt gtttatgttg agtgatgcag
     1201 acttagaaca ttttaaatta cataaaggta atttttgcgg taatagattt tgtccaatgt
     1261 gtagttggcg acttgcttgt aaggatagtt tagaaatatc tattcttatg gagcatttaa
     1321 gaaaagaaga aaataaagag tttatatttt taactcttac aactccaaat gtaaaaagtt
     1381 atgatcttaa ttattctatt aaacaatata ataaatcttt taaaaaatta atggagcgta
     1441 aggaagttaa ggatataact aaaggttata taagaaaatt agaagtaact taccaaaagg
     1501 aaaaatacat aacaaaggat ttatggaaaa taaaaaaaga ttattatcaa aaaaaaggac
     1561 ttgaaattgg tgatttagaa cctaattttg atacttataa tcctcatttt catgtagtta
     1621 ttgcagttaa taaaagttat tttacagata aaaattatta tataaatcga gaaagatggt
     1681 tggaattatg gaagtttgct actaaggatg attctataac tcaagttgat gttagaaaag
     1741 caaaaattaa tgattataaa gaggtttacg aacttgcgaa atattcagct aaagacactg
     1801 attatttaat atcgaggcca gtatttgaaa ttttttataa agcattaaaa ggcaagcagg
     1861 tattagtttt tagtggattt tttaaagatg cacacaaatt gtacaagcaa ggaaaacttg
     1921 atgtttataa aaagaaagat gaaattaaat atgtctatat agtttattat aattggtgca
     1981 aaaaacaata tgaaaaaact agaataaggg aacttacgga agatgaaaaa gaagaattaa
     2041 atcaagattt aatagatgaa atagaaatag attaaagtgt aactatactt tatatatata
     2101 tgattaaaaa aataaaaaac aacagcctat taggttgttg ttttttattt tctttattaa
     2161 tttttttaat ttttagtttt tagttctttt ttaaaataag tttcagcctc tttttcaata
     2221 ttttttaaag aaggagtatt tgcatgaatt gccttttttc taacagactt aggaaatatt
     2281 ttaacagtat cttcttgcgc cggtgatttt ggaacttcat aacttactaa tttataatta
     2341 ttattttctt ttttaattgt aacagttgca aaagaagctg aacctgttcc ttcaactagt
     2401 ttatcatctt caatataata ttcttgacct atatagtata aatatatttt tattatattt
     2461 ttactttttt ctgaatctat tattttataa tcataaaaag ttttaccacc aaaagaaggt
     2521 tgtactcctt ctggtccaac atattttttt actatattat ctaaataatt tttgggaact
     2581 ggtgttgtaa tttgattaat cgaacaacca gttatactta aaggaattat aactataaaa
     2641 atatatagga ttatcttttt aaatttcatt attggcctcc tttttattaa atttatgtta
     2701 ccataaaaag gacataacgg gaatatgtag aatattttta atgtagacaa aattttacat
     2761 aaatataaag aaaggaagtg tttgtttaaa ttttatagca aactatcaaa aattaggggg
     2821 ataaaaattt atgaaaaaaa ggttttcgat gttattttta tgtttaactt taatagtttg
     2881 tggtttattt acaaattcgg ccggcccgct tgagaccacc ccaggataca tagattacca
     2941 caactccgag cccttccacc tagcgtgcta accacgcacg ctattgttgt atatcccaat
     3001 ggcgcgccga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     3061 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     3121 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3181 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3241 gggcgctgtt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3301 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3361 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3421 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3481 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3541 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3601 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3661 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3721 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3781 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3841 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3901 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3961 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4021 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4081 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4141 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4201 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4261 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4321 ataccgcgag aaccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4381 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4441 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4501 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4561 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4621 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4681 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4741 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4801 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4861 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4921 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4981 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5041 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5101 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5161 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     5221 aataggcgta tcacgaggcc cttttgtc
//
