LOCUS       sequence-362791-        5507 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5507)
  AUTHORS   Le Y, Miller JL, Sauer B
  TITLE     GFPcre fusion vectors with enhanced expression.
  JOURNAL   Anal Biochem. 1999 Jun 1. 270(2):334-6.
  PUBMED    10334853
REFERENCE   2  (bases 1 to 5507)
  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: "Anal 
            Biochem. 1999 Jun 1. 270(2):334-6."
COMMENT     Sequence Label: pBS595 tet-hCMV-EGFPcre
FEATURES             Location/Qualifiers
     source          1..5507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     110..129
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     323..345
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     337..354
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     338..354
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    378..396
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    420..438
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    462..480
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    504..522
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    546..564
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    588..606
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        639..677
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     674..698
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      807..816
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(813..815,816..818,819..1529)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(813..815,816..818,819..1529)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1119..1138)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1466..1487
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1584..2615
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MANLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     primer_bind     complement(3245..3261)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3245..3261)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3258..3280)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3269..3285
                     /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(3293..3299,3300..3317,3318..3323))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3338..3359
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3476..3493)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3647..4235)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3727..3746)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4406..5197,5198..5266))
                     /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     5029..5048
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5267..5371)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5439..5457
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(5495..5507,1..10))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
       61 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
      121 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
      181 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
      241 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
      301 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag
      361 cttctcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact
      421 ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga
      481 aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagttt
      541 accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga
      601 tagagaaaag tgaaagtcga gctcggtacc cgggtcgagg taggcgtgta cggtgggagg
      661 cctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct
      721 gttttgacct ccatagaaga caccgggacc gatccagcct ccgcggcccc gaattcgagc
      781 tcggtacccg gggatctgca ccggtcgcca ccatggtgag caagggcgag gagctgttca
      841 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      901 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
      961 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     1021 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1081 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1141 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1201 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1261 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     1321 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1381 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     1441 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1501 tcactctcgg catggacgag ctgtacaagt ccggtacagc tctcgacgga gaaagctcag
     1561 gctctggctc agagtctgac tccatggcca atttactgac cgtacaccaa aatttgcctg
     1621 cattaccggt cgatgcaacg agtgatgagg ttcgcaagaa cctgatggac atgttcaggg
     1681 atcgccaggc gttttctgag catacctgga aaatgcttct gtccgtttgc cggtcgtggg
     1741 cggcatggtg caagttgaat aaccggaaat ggtttcccgc agaacctgaa gatgttcgcg
     1801 attatcttct atatcttcag gcgcgcggtc tggcagtaaa aactatccag caacatttgg
     1861 gccagctaaa catgcttcat cgtcggtccg ggctgccacg accaagtgac agcaatgctg
     1921 tttcactggt tatgcggcgg atccgaaaag aaaacgttga tgccggtgaa cgtgcaaaac
     1981 aggctctagc gttcgaacgc actgatttcg accaggttcg ttcactcatg gaaaatagcg
     2041 atcgctgcca ggatatacgt aatctggcat ttctggggat tgcttataac accctgttac
     2101 gtatagccga aattgccagg atcagggtta aagatatctc acgtactgac ggtgggagaa
     2161 tgttaatcca tattggcaga acgaaaacgc tggttagcac cgcaggtgta gagaaggcac
     2221 ttagcctggg ggtaactaaa ctggtcgagc gatggatttc cgtctctggt gtagctgatg
     2281 atccgaataa ctacctgttt tgccgggtca gaaaaaatgg tgttgccgcg ccatctgcca
     2341 ccagccagct atcaactcgc gccctggaag ggatttttga agcaactcat cgattgattt
     2401 acggcgctaa ggatgactct ggtcagagat acctggcctg gtctggacac agtgcccgtg
     2461 tcggagccgc gcgagatatg gcccgcgctg gagtttcaat accggagatc atgcaagctg
     2521 gtggctggac caatgtaaat attgtcatga actatatccg taacctggat agtgaaacag
     2581 gggcaatggt gcgcctgctg gaagatggcg attagccatt aacgcgtaaa tgattgctct
     2641 agagtgaggg tccccacctg ggacccttga gagtatcagg tctcccacgt gggagacaag
     2701 aaatccctgt ttaatattta aacagcagtg ttccccatct gggtccttgc acccctcact
     2761 ctggcctcag ccgactgcac agcggcccct gcatcccctt ggctgtgagg cccctggaca
     2821 agcagaggtg gccagagctg ggaggcatgg ccctggggtc ccacgaattt gctggggaat
     2881 ctcgtttttc ttcttaagac ttttgggaca tggtttgact cccgaacatc accgacgtgt
     2941 ctcctgtttt tctgggtggc ctcgggacac ctgccctgcc cccacgaggg tcaggactgt
     3001 gactcttttt agggccaggc aggtgcctgg acatttgcct tgctggacgg ggactgggga
     3061 tgtgggaggg agcagacagg aggaatcatg tcaggcctgt gtgtgaaagg aagctccact
     3121 gtcaccctcc acctcttcac cccccactca ccagtgtccc ctccactgtc acattgtaac
     3181 tgaacttcag gataataaag tgtttgcctc caaaaaaaaa aaaaaaaacc gaattcgtaa
     3241 tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     3301 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     3361 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     3421 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     3481 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3541 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3601 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3661 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3721 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3781 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3841 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3901 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3961 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4021 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4081 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4141 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4201 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4261 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4321 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4381 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4441 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4501 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4561 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4621 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4681 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4741 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4801 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4861 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4921 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4981 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5041 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5101 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5161 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5221 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5281 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5341 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5401 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     5461 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatg
//
