LOCUS       sequence-439438-        7132 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescently tagged Sep11-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7132)
  TITLE     Ori lab plasmids
REFERENCE   2  (bases 1 to 7132)
  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: Sep11-GFP
FEATURES             Location/Qualifiers
     source          1..7132
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(499..523)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    533..551
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    554..572
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(574..777)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(603..623)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        778..1157
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1329..1348
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1423..1527
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1528..1596,1597..2388)
                     /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     complement(1746..1765)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2559..3147
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3048..3067
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3301..3318
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3435..3456
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3471..3476,3477..3494,3495..3501)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3509..3525
                     /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)."
     primer_bind     3514..3536
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3533..3549
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3533..3549
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3598..3719
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3609..3628)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3663..3682
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3849..4871)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(3849..4871)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(4879..4926)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     5058..5077
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    5210..5434
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5423..5440
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(5484..6197,6198..6200,6201..6203))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     5878..5897
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6137..6158
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(6216..6236)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 tcaattcctc ttgcaggtag gcctcgaact gggcatcaat atattctact atcggcttat
       61 agctgtcatc tttatttatc tggtctccaa atcccacggt gtcaacaatg gttaacttca
      121 gccgtacatt gctttcctga agctcataac ttctggcttt taaccgaaca cctggttcat
      181 tgtgagtagc tgggtcactt tcaaatttgg tgttgaacaa agtgtccatt aacgtggatt
      241 tgccaatgcc tgtctcacca acacaaagga tgttgaaaca gaatccttga gaagtagact
      301 tgttgaccag ctggtcaggg aggctgtcaa atcccacatg gccagacaaa gacaagtttc
      361 gaagctcttc attagacggt ctccccacgg ccacggccat gcaggcctaa gcttaagttt
      421 aaacgctaga gtccggaggc tggatcggtc ccggtgtctt ctatggaggt caaaacagcg
      481 tggatggcgt ctccaggcga tctgacggtt cactaaacga gctcgtcgac gatctctatc
      541 actgataggg agatctctat cactgatagg gagagctctg cttatataga cctcccaccg
      601 tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc
      661 ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa
      721 tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc gcatcaccat
      781 ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat
      841 gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact
      901 tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca
      961 ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt
     1021 caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac
     1081 gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact attaataact
     1141 agtcaataat caatgtcaac gcgtatatct ggcccgtaca tcgcgaagca gcgcaaaacg
     1201 cctaacccta agcagattct tcatgcaatt gtcggtcaag ccttgccttg ttgtagctta
     1261 aattttgctc gcgcactact cagcgacctc caacacacaa gcagggagca gatactggct
     1321 taactatgcg gcatcagagc agattgtact gagagtgcac cataggggat cgggagatct
     1381 cccgatccgt cgacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     1441 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1501 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     1561 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     1621 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     1681 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     1741 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     1801 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     1861 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     1921 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     1981 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     2041 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     2101 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     2161 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     2221 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     2281 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     2341 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     2401 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     2461 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2521 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2581 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2641 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2701 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2761 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2821 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2881 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2941 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3001 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3061 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3121 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3181 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     3241 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     3301 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     3361 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     3421 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     3481 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     3541 agctatgacc atgattacgc caagctctag ctagaggtcg acggtataca gacatgataa
     3601 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     3661 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagttg
     3721 gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc
     3781 cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt
     3841 agtacgtgct attcctttgc cctcggacga gtgctggggc gtcggtttcc actatcggcg
     3901 agtacttcta cacagccatc ggtccagacg gccgcgcttc tgcgggcgat ttgtgtacgc
     3961 ccgacagtcc cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca
     4021 tcatcgaaat tgccgtcaac caagctctga tagagttggt caagaccaat gcggagcata
     4081 tacgcccgga gccgcggcga tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc
     4141 tgctgctcca tacaagccaa ccacggcctc cagaagaaga tgttggcgac ctcgtattgg
     4201 gaatccccga acatcgcctc gctccagtca atgaccgctg ttatgcggcc attgtccgtc
     4261 aggacattgt tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg
     4321 gcccaaagca tcagctcatc gagagcctgc gcgacggacg cactgacggt gtcgtccatc
     4381 acagtttgcc agtgatacac atggggatca gcaatcgcgc atatgaaatc acgccatgta
     4441 gtgtattgac cgattccttg cggtccgaat gggccgaacc cgctcgtctg gctaagatcg
     4501 gccgcagcga tcgcatccat ggcctccgcg accggctgca gaacagcggg cagttcggtt
     4561 tcaggcaggt cttgcaacgt gacaccctgt gcacggcggg agatgcaata ggtcaggctc
     4621 tcgctgaatt ccccaatgtc aagcacttcc ggaatcggga gcgcggccga tgcaaagtgc
     4681 cgataaacat aacgatcttt gtagaaacca tcggcgcagc tatttacccg caggacatat
     4741 ccacgccctc ctacatcgaa gctgaaagca cgagattctt cgccctccga gagctgcatc
     4801 aggtcggaga cgctgtcgaa cttttcgatc agaaacttct cgacagacgt cgcggtgagt
     4861 tcaggctttt tggccaagga agttcctata ctttctagag aataggaact tcggaatagg
     4921 aacttctagg tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     4981 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     5041 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     5101 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     5161 gcgtggggat accccctaga gccccagctg gttctttccg cctcagaagc catagagccc
     5221 accgcatccc cagcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc
     5281 accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt
     5341 tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg
     5401 caaacaacag atggctggca actagaaggc acagtcgagg ctgatcagcg ggtttaaacg
     5461 ggccctctag actcgagtca ctattacttg tacagctcgt ccatgccgag agtgatcccg
     5521 gcggcggtca cgaactcctt aaggaccatg tgatcgcgct tctcgttggg gtctttgctc
     5581 agggcggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg
     5641 ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg
     5701 atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg
     5761 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
     5821 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
     5881 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
     5941 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
     6001 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     6061 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
     6121 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     6181 agctcctcgc ccttgctcac catggtggta ccttcacctt gaaaatataa attttcgcgg
     6241 ccgcgtgtga agcttgcatt tttcttatcc ttgtctttct tggtttgctg ggccccagat
     6301 tgctgggcct gggactgtag taactgagcc gctgctttct tcttctggaa gttgttcacc
     6361 tcctcctcaa gctccttctt cttgtcttcc actttcttct tttcttcttg gtgtgtccgc
     6421 tttagaaggt caaacttctc gtgaagctct ttctctgcct ccttaagttc agcttctttc
     6481 tccttcactc tcataacaaa catttgtctc atttcttctt ctttcttctg cagttctccc
     6541 aggaattcat tcctttttgc ttcatatgtc tcctgaagac tgaagggttt gctgtcaggg
     6601 tcagtgtcct tgaaccccat ctcttcaagc ttacagcgtc ggtacaattc atagtggcgg
     6661 gtgtgagtct gctctcgcaa gtcctccatg ttcacgcgga tcagcatctc tcgaagtttc
     6721 acaaaatcgc aatgattttc attctcaacc tgcaccacac cccaggggta ctgcctggcc
     6781 tttgccatct tgttgccaat cttcacctct tcggtgctgc caaccactgc aaatgggaga
     6841 tggacactca ttgttgcgtt aatctctgcc accgtttctt catcagtggg aaactgatat
     6901 atctggaccc cattgctgac cagttcactc atgatcttac tcttgaattt gtgcagttca
     6961 ttcttggcaa tggtgtcagc ttttgcaatt attggaatga tgttcacctt actgtccagc
     7021 tttttcatgg tgaccagatc cagggacttt agtgaatgtc cagtaggggc aataaagtag
     7081 aggcaggcat ggatcctcgt gtcatggtag ttgaagagag aacgtttaat ct
//
