LOCUS       sequence-163454-        7927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cre under the control of the elongation factor-1 alpha 
            (EF1alpha) promoter; includes upstream Kozak modification to 
            increase expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7927)
  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 7927)
  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: pBS513 EF1alpha-cre
FEATURES             Location/Qualifiers
     source          1..7927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..1032
                     /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="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     RBS             2684..2692
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(3486..3503)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3486..3502)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3495..3517)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3715..4170
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3802..3821)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4012..4033
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4187..4206)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4306..4328
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4366..4384)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4452..4556
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4557..4625,4626..5417)
                     /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(4775..4794)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5588..6176
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6077..6096
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6330..6347
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6464..6485
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6500..6505,6506..6523,6524..6530)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6538..6554
                     /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     6543..6565
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6562..6578
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6562..6578
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6677..7855
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6908..7846
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7803..7823
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 atgtccaatt tactgaccgt acaccaaaat ttgcctgcat taccggtcga tgcaacgagt
       61 gatgaggttc gcaagaacct gatggacatg ttcagggatc gccaggcgtt ttctgagcat
      121 acctggaaaa tgcttctgtc cgtttgccgg tcgtgggcgg catggtgcaa gttgaataac
      181 cggaaatggt ttcccgcaga acctgaagat gttcgcgatt atcttctata tcttcaggcg
      241 cgcggtctgg cagtaaaaac tatccagcaa catttgggcc agctaaacat gcttcatcgt
      301 cggtccgggc tgccacgacc aagtgacagc aatgctgttt cactggttat gcggcggatc
      361 cgaaaagaaa acgttgatgc cggtgaacgt gcaaaacagg ctctagcgtt cgaacgcact
      421 gatttcgacc aggttcgttc actcatggaa aatagcgatc gctgccagga tatacgtaat
      481 ctggcatttc tggggattgc ttataacacc ctgttacgta tagccgaaat tgccaggatc
      541 agggttaaag atatctcacg tactgacggt gggagaatgt taatccatat tggcagaacg
      601 aaaacgctgg ttagcaccgc aggtgtagag aaggcactta gcctgggggt aactaaactg
      661 gtcgagcgat ggatttccgt ctctggtgta gctgatgatc cgaataacta cctgttttgc
      721 cgggtcagaa aaaatggtgt tgccgcgcca tctgccacca gccagctatc aactcgcgcc
      781 ctggaaggga tttttgaagc aactcatcga ttgatttacg gcgctaagga tgactctggt
      841 cagagatacc tggcctggtc tggacacagt gcccgtgtcg gagccgcgcg agatatggcc
      901 cgcgctggag tttcaatacc ggagatcatg caagctggtg gctggaccaa tgtaaatatt
      961 gtcatgaact atatccgtaa cctggatagt gaaacagggg caatggtgcg cctgctggaa
     1021 gatggcgatt agccattaac gcgtaaatga ttgctataat tatttgatat ttatggtgac
     1081 atatgagaaa ggatttcaac atcgacggaa aatatgtagt gctgtctgta agcactaata
     1141 ttcagtcgcc agccgtcatt gtcactgtaa agctgagcga tagaatgcct gatattgact
     1201 caatatccgt tgcgtttcct gtcaaaagta tgcgtagtgc tgaacatttc gtgatgaatg
     1261 ccaccgagga agaagcacgg cgcggttttg ctaaagtgat gtctgagttt ggcgaactct
     1321 tgggtaaggt tggaattgtc gagggatctc ggaatggacc ccaactgctc ctgctccacc
     1381 ggtaagactc ccgatccttg gtctttagaa taccaagttg ggaccgcaga gcggaatccc
     1441 cgagttgtag aggcttggcg ggaataggca cctttagttg gcgattcatt ccggttcttt
     1501 ctagaatccg ctcttgcaaa agccttcatt agttacgagt attgtcgaac gggtcctttg
     1561 gcggggttgg ggctaggatt tagacgcgca aatgtccggt tcctgatcac ccagttagtg
     1621 gggacatctg ggttgagtcc caggcattac taaacttact gtgaattgct tgaattaaga
     1681 aagaggtgaa ggacctttat gtcttgggac tcaaagacat aatccctgac ttaacctgtg
     1741 aggagaaaag tggggctagg ctccctgcag ctccgaggag gacttagtga actgagccgg
     1801 gactcgtggt gttggccact gctgtaatgc tgcctccctc atgctgtctt ctttctcctc
     1861 cccaggcggc tcctgcactt gcaccagctc ctgcgcctgc aagaactgca agtgcacctc
     1921 ctgcaagaag agtgagttgg gacaccttgg gtggcggcta aggctagggg cggggaactc
     1981 ctacaaaact ggctctgaga aatgtccttt gcttcccgga ggccattgta ttgtctcggg
     2041 gacagaacta tacagagaac tatttaaaaa aaccgaggtc ttctctgttg gggacaggaa
     2101 gcagaggtct tcagccaggc tgacctcttc ctcctccttt ctaggctgct gctcctgctg
     2161 tcccgtgggc tgctccaaat gtgcccaggg ctgtgtctgc aaaggcgccg cggacaagtg
     2221 cacgtgctgt gcctgatgtg acgaacagcg ctgccaccac gtgtaaatag tatcggacca
     2281 acccagcgtc ttcctataca gttccaccct gtttactaaa cccccgtttt ctaccgagta
     2341 cgtgaataat aaaagcctgt ttgagtctaa ctctggtttt cttggtgtgg tttggcaata
     2401 agaaactggg gtgacttgat agtctgggga tctggttttg gaccccctcg tgcctttacc
     2461 tcccgcctct ggccctcaca gaggggtaat gtctttgggt aaagccaagc tatcccataa
     2521 gcttcctcat ggaaaacagc tgtgtgcctg gtcaggtctt gtgttagggg aagggaggga
     2581 gggagggtgg tcctggaaga cttgatgtcc catcctggaa gcacagaaaa tcttatcatt
     2641 tatattctct ttggattctg gtttcctctc acattgttgg gtataaggag gtgggtatga
     2701 aaattaacat agggtgtctc gtatgctagg cagggactaa cactgaacta gattccagag
     2761 gggctttctt ggctgttttg acacaagttg cttaccttgt cctagaactc attctgcagt
     2821 ccagtctgac cttgagtttt tgattctcct gcctcagctt cccaagtacc tgggattaca
     2881 ggagtgactg tgtcaccagg ccatctccta ttcagccacc aaatgcagac actattgcat
     2941 atgccatcaa gattttgctg aaaggaccct gataaagctg tctcctgtga ggctttgcca
     3001 gtgcccagca aacacagaag tggatgctca cagtcagcta ttggatggaa cacagggccc
     3061 ccaatggagg agctagagaa agtacccaag gagctaaagg gatctgcaac cctataggtg
     3121 gaacagcaat ctgaactaac cagtaccccc tgaactcgtg tatctagctt cgtatgtagc
     3181 agaagatggc ctagttggcc atcactggga agagaggacc cttggtctag caaactatat
     3241 gccccagttc aggggaaagc cagggccaag aagtgggagt gagtgggtag gggagcgggg
     3301 tgggggtagg ggtggggtat aggggacttt tgggatagca tttgaaatat aaacgaagaa
     3361 aatatccaaa aaaaaaaaga tggaaaaaaa agtgataaag agcgtgggtc ctgcctgtaa
     3421 tcaagatatt tgagatgtag cagcaggaag gccaggaatt ggtcgagggt accgagctcg
     3481 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     3541 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     3601 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
     3661 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     3721 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3781 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3841 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3901 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     3961 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4021 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     4081 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4141 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     4201 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     4261 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     4321 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     4381 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     4441 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4501 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4561 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4621 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4681 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4741 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4801 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4861 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4921 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4981 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5041 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5101 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5161 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5221 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5281 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5341 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5401 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     5461 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5521 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5581 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5641 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5701 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5761 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5821 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5881 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5941 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6001 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6061 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6121 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6181 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6241 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     6301 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     6361 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     6421 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     6481 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     6541 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctttgca
     6601 aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct tctaggtctt
     6661 gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     6721 ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg
     6781 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     6841 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     6901 aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc
     6961 ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc ccgagcttcg
     7021 ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc
     7081 ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg
     7141 cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc
     7201 gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat
     7261 ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc
     7321 gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg
     7381 gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct
     7441 ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg
     7501 gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag
     7561 gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc
     7621 gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga
     7681 ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc
     7741 ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat
     7801 tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaggaat
     7861 tggatccaga ttcgaggggc agagccgatc ctgtacactt tacttaaaac cattatctga
     7921 gtgtgaa
//
