LOCUS       sequence-350203-        4929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Helper vector for the transient expression of Cre recombinase in 
            filamentous fungi. Not replicative in fungi..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4929)
  AUTHORS   Roux I, Chooi YH
  TITLE     Cre/lox-Mediated Chromosomal Integration of Biosynthetic Gene 
            Clusters for Heterologous Expression in Aspergillus nidulans.
  JOURNAL   ACS Synth Biol. 2022 Feb 16. doi: 10.1021/acssynbio.1c00458.
  PUBMED    35168324
REFERENCE   2  (bases 1 to 4929)
  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: "ACS Synth 
            Biol. 2022 Feb 16. doi: 10.1021/acssynbio.1c00458."
COMMENT     Sequence Label: pGem-PgpdA-Cre-TtrpC
FEATURES             Location/Qualifiers
     source          1..4929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..47)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(201..789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(281..300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(960..1751,1752..1820))
                     /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     1583..1602
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1821..1925)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2003..2458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2140..2161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2352..2371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2584..2606
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2598..2615
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2599..2615
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3352..3368)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3353..3369)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4728..4744)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4728..4744)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4741..4763)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4752..4768
                     /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(4776..4782,4783..4800,4801..4806))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4821..4842
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
       61 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
      121 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      181 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      241 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      301 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      361 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      421 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      481 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      541 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
      601 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
      661 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      721 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
      781 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
      841 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
      901 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
      961 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     1021 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     1081 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     1141 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     1201 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     1261 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     1321 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     1381 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     1441 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     1501 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     1561 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     1621 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     1681 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     1741 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     1801 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     1861 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     1921 ccgcgcacat ttccccgaaa agtgccacct gatgcggtgt gaaataccgc acagatgcgt
     1981 aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta
     2041 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     2101 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     2161 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     2221 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     2281 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     2341 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     2401 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     2461 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     2521 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     2581 tttcccagtc acgacgttgt aaaacgacgg ccagtgaaca tgcggagaga cggacggacg
     2641 cagagagaag ggctgagtaa taagcgccac tgcgccagac agctctggcg gctctgaggt
     2701 gcagtggatg attattaatc cgggaccggc cgcccctccg ccccgaagtg gaaaggctgg
     2761 tgtgcccctc gttgaccaag aatctattgc atcatcggag aatatggagc ttcatcgaat
     2821 caccggcagt aagcgaagga gaatgtgaag ccaggggtgt atagccgtcg gcgaaatagc
     2881 atgccattaa cctaggtaca gaagtccaat tgcttccgat ctggtaaaag attcacgaga
     2941 tagtaccttc tccgaagtag gtagagcgag tacccggcgc gtaagctccc taattggccc
     3001 atccggcatc tgtagggcgt ccaaatatcg tgcctctcct gctttgcccg gtgtatgaaa
     3061 ccggaaaggc cgctcaggag ctggccagcg gcgcagaccg ggaacacaag ctggcagtcg
     3121 acccatccgg tgctctgcac tcgacctgct gaggtccctc agtccctggt aggcagcttt
     3181 gccccgtctg tccgcccggt gtgtcggcgg ggttgacaag gtcgttgcgt cagtccaaca
     3241 tttgttgcca tattttcctg ctctccccac cagctgctct tttcttttct ctttcttttc
     3301 ccatcttcag tatattcatc ttcccatcca agaaccttta atcaagctta tcgataccgt
     3361 cgacctcgac tctagaggat cactcgtcgt gtcgcccatg tctaatttgt tgactgttca
     3421 tcaaaatttg ccagctttgc cagttgatgc aacgtctgat gaggtaagaa agaatttgat
     3481 ggatatgttt agagatagac aagcattctc tgaacatact tggaaaatgt tgttgtctgt
     3541 ttgtagatct tgggctgctt ggtgtaaatt aaataataga aaatggtttc cagctgaacc
     3601 agaagatgtt agagattatt tgttgtattt gcaagctaga ggtttggctg ttaaaactat
     3661 acaacaacat ttgggtcaat taaatatgtt gcataggaga tctggtttgc ctagaccatc
     3721 tgattctaat gctgtatctt tggttatgag aaggattaga aaagaaaatg ttgatgcagg
     3781 tgaaagagct aaacaagcat tggcgtttga aagaactgat tttgatcaag ttagatcttt
     3841 gatggaaaat tctgatagat gtcaagatat aagaaacttg gctttcttgg gaattgctta
     3901 taatacattg ttgagaattg ctgaaattgc tagaattaga gttaaagaca tttctagaac
     3961 tgatggtggt agaatgttga ttcatattgg tagaactaaa acattggttt caactgctgg
     4021 cgttgagaaa gcattgtctt tgggtgttac taaattggtt gaaagatgga tttctgtatc
     4081 tggtgtggct gatgacccaa ataattattt attttgtaga gttagaaaga atggtgttgc
     4141 ggctccatct gctacatctc agctctctac ccgtgccttg gaaggaattt ttgaagcaac
     4201 tcatagattg atttatggtg ctaaagatga ttctggtcaa agatatttag catggtctgg
     4261 tcattctgct agagttggtg ccgctagaga tatggctaga gctggggtat ctattcctga
     4321 aattatgcaa gctggtggtt ggactaatgt taacatcgtc atgaactata ttagaaattt
     4381 ggattctgaa actggtgcaa tggttagact cttggaagat ggtgattagg gcgacgacga
     4441 cgttcgaggg ggggcccggt accgccccgt ccggtcctgc ccgtcaccga gatccactta
     4501 acgttactga aatcatcaaa cagcttgacg aatctggata taagatcgtt ggtgtcgatg
     4561 tcagctccgg agttgagaca aatggtgttc aggatctcga taagatacgt tcatttgtcc
     4621 aagcagcaaa gagtgccttc tagtgattta atagctccat gtcaacaaga ataaaacgcg
     4681 tttcgggttt acctcttcca gatacagctc atctgcaatg catcatggtc atagctgttt
     4741 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4801 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4861 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4921 gggagaggc
//
