LOCUS       sequence-176987-        6189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pET LIC cloning vector for yORF-tdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6189)
  TITLE     C-terminal fluorescent fusion vectors for E. coli expression
REFERENCE   2  (bases 1 to 6189)
  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"
COMMENT     Sequence Label: pET tdTomato LIC cloning vector (u-tdTomato)
FEATURES             Location/Qualifiers
     source          1..6189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..46
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(126..143)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(297..885)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(377..396)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1056..1847,1848..1916))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1679..1698
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1917..2021)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2089..2107
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(2134..2139,2140..2156,2157..2162)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(2528..2575)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2571..2589
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             4178..4200
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4288..4307)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4289..4307)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4391..4410)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             5868..6059
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6053..6075)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    join(6161..6189,1..111)
                     /label=bom
                     /note="basis of mobility region from pBR322"
ORIGIN
        1 gcggagtgta tactggctta actatgcggc atcagagcag attgtactga gagtgcacca
       61 ttgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc
      121 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      181 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      241 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      301 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      361 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      421 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      481 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      541 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      601 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      661 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      721 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      781 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      841 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      901 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      961 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     1021 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1081 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1141 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     1201 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1261 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     1321 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc
     1381 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1441 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1501 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1561 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1621 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat
     1681 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1741 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1801 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1861 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     1921 ttcctttttc aatgttattg aagcatttat cagggttatt gtctcatgag cggatacata
     1981 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     2041 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     2101 acgaggccct ttcgtcttca agaattctca tgtttgacag cttatcatcg ataagcttta
     2161 atgcggtagt ttatcacagt taaattgcta acgcagtcag gcaccgtgta tgaaatctaa
     2221 caatgcgctc atcgtcatcc tcggcaccgt caccctggat gctgtaggca taggcttggt
     2281 tatgccggta ctgccgggcc tcttgcggga tgtcgtccat tccgacagca tcgccagtca
     2341 ctatggcgtg ctgctagcgc tatatgcgtt gatgcaattt ctatgcgcac ccgttctcgg
     2401 agcactgtcc gaccgctttg gccgccgccc agtcctgctc gcttcgctac ttggagccac
     2461 tatcgactac gcgatcatgg cgaccacacc cgtcctgtgg atgtccggat atagttcctc
     2521 ctttcagcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat gctagttatt
     2581 gctcagcggt ggcagcagcc aactcagctt cctttcgggc tttgttagca gccggatcag
     2641 cgcgcacgcg tggtaccggc cggccaccag gtggcgcgcc gcgatcgcgg atccttatgg
     2701 agttggttat tacttataca gctcatccat gccgtacaga aacagatggt ggcgtccttc
     2761 agacctctca tactgttcca caatggtgta gtcttcattg tgggatgtga tatccagctt
     2821 ggtgtccaca tagtaatagc cgggcagctg cactggtttc ttggccatgt agatggtttt
     2881 gaactccacc aggtagtggc cgccgtcttt cagcttcagg gcctggtgga tttcgccctt
     2941 cagcactccg tctctaggat acagccgttc agtgctggcc tcccacccca ttgttttctt
     3001 ctgcatcacg ggcccatcgg gtggaaaatt ggtgccgcgc attttgactt tgtagatcag
     3061 agtgccatcc tgcagagagg agtcctgtgt cacggtcacc aggcctccat cttcaaaatt
     3121 catcactctc tcccacttaa agccctctgg gaaagacagt ttcttgtaat caggaatgtc
     3181 agcgggatgc ttgacatagg ccttacttcc gtacataaac tgaggagaca ggatatccca
     3241 agcgaaaggc agtggtcctc ccttggtcac tttcagcttg gcagtctgtg tgccctcgta
     3301 aggcctccct tcgccctctc cttcaatctc aaattcgtgc ccattcatgc tgccctccat
     3361 gcgcacctta aacctcatga actctttgat cacagccata ttgttatcct cgcttgaggc
     3421 ggttccacta gaccctgagc cggtacttcc agtcccatgg cccaggaaca gatggtgtct
     3481 cccctcgctc cgttcatact gctccacaat agtgtagtcc tcgttgtgtg aagtgatatc
     3541 cagttttgtg tccacgtaat agtagcctgg cagctgcaca ggtttcttgg ccatgtagat
     3601 tgtcttgaac tccaccaggt aatgtccccc atctttcagc ttcagagcct ggtggatttc
     3661 tcctttcagc accccgtcgc gtggatacag cctttcggtg gaggcctccc atcccatagt
     3721 tttcttctgc atcacaggtc catcaggggg aaaatttgtc cctctcattt tcaccttgta
     3781 gatcagggtc ccatcctgca gggagctgtc ctgggtcaca gtcaccaggc ctccgtcctc
     3841 gaagttcatc actctttccc acttaaaccc ctcggggaag ctcagtttct tataatcggg
     3901 aatgtcagct ggatgtttca cataggcctt ggagccgtac ataaactgtg ggctcaggat
     3961 gtcccaagcg aatggcagag gcccgccctt tgtcactttc agcttggctg tctgggtgcc
     4021 ctcgtagggc cgtccctctc cttctccctc aatttcgaac tcgtggccgt tcatgcttcc
     4081 ttccattctc accttaaacc gcatgaactc tttgatcact tcctcgccct tactcacgga
     4141 tccgctgctg ccgttggagg atgagaggat atctatatct ccttcttaaa gttaaacaaa
     4201 attatttcta gaactcgagt cctgcagggc ggccgcttaa ttaaagagct ctccggagcc
     4261 ggcactagag ggaaaccgtt gtggtctccc tatagtgagt cgtattaatt tcgcgggatc
     4321 gagatctcga tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg
     4381 gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg
     4441 ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg
     4501 ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta
     4561 ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgacc gatgcccttg
     4621 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca
     4681 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc
     4741 attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta
     4801 ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc
     4861 ggcgagaagc aggccattat cgccggcatg gcggccgacg cgctgggcta cgtcttgctg
     4921 gcgttcgcga cgcgaggctg gatggccttc cccattatga ttcttctcgc ttccggcggc
     4981 atcgggatgc ccgcgttgca ggccatgctg tccaggcagg tagatgacga ccatcaggga
     5041 cagcttcaag gatcgctcgc ggctcttacc agcctaactt cgatcattgg accgctgatc
     5101 gtcacggcga tttatgccgc ctcggcgagc acatggaacg ggttggcatg gattgtaggc
     5161 gccgccctat accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc
     5221 tcgacctgaa tggaagccgg cggcacctcg ctaacggatt caccactcca agaattggag
     5281 ccaatcaatt cttgcggaga actgtgaatg cgcaaaccaa cccttggcag aacatatcca
     5341 tcgcgtccgc catctccagc agccgcacgc ggcgcatctc gggcagcgtt gggtcctggc
     5401 cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc
     5461 ttactggtta gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca
     5521 aaacgtctgc gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc
     5581 tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct
     5641 gctggctacc ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag
     5701 tgatttttct ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca
     5761 gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg
     5821 gtatcattac ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg
     5881 aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga
     5941 aactcaacga gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg
     6001 ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac
     6061 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     6121 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac
     6181 gtagcgata
//
