LOCUS       sequence-308575-        5327 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Creb1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5327)
  TITLE     Creb1  plasmid
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5327)
  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: 
            "Unpublished"
COMMENT     Sequence Label: Creb1
FEATURES             Location/Qualifiers
     source          1..5327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        554..572
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             616..639
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1690..1706)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1691..1707)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1762..1781)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1763..1781)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2055..2176
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2092..2111)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2146..2165
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2183..2638)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2320..2341)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2532..2551
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2665..2769
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2771..3128
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2771..2962
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2794..2814)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2979..3114
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3163..3957
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3163..3957
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3217..3236)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3827..3846
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4145..4164)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4189..4236
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4565..5153
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5054..5073
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgatt
      541 acgccaagct cgaaattaac cctcactaaa gggaacaaaa gctggagctc caccgcggtg
      601 gcggccgcca ccatggatta caaggatgac gacgataagg cccgggcgga tcccccgggc
      661 tgcaggaatt cgattatgac catggaatct ggagcagaca accagcagag tggagatgct
      721 gctgtaacag aagctgaaaa tcaacaaatg acagttcaag cccagccaca gattgccaca
      781 ttagcccagg tatccatgcc agcagctcat gcaacatcat ctgctcccac tgtaacctta
      841 gtgcagctgc ccaatgggca gacagtccag gtccatggcg ttatccaggc ggcccagcca
      901 tcagttatcc agtctccaca agtccaaaca gttcagattt caactattgc agaaagtgaa
      961 gattcacagg agtctgtgga tagtgtaact gattcccaaa aacgaaggga aatcctttca
     1021 aggaggcctt cctacaggaa aattttgaat gacttatctt ctgatgcacc aggggtgcca
     1081 aggattgaag aagaaaagtc agaagaggag acttcagccc ctgccatcac cactgtaaca
     1141 gtgccaaccc ccatttacca aactagcagt gggcagtaca ttgccattac ccagggagga
     1201 gcaatacagc tggctaacaa tggtacggat ggggtacagg gcctgcagac attaaccatg
     1261 accaatgcag ctgccactca gccgggtact accattctac agtatgcaca gaccactgat
     1321 ggacagcaga ttctagtgcc cagcaaccaa gttgttgttc aagctgcctc aggcgatgta
     1381 caaacatacc agatccgcac agcacccacg agcaccattg cccctggagt tgttatggcg
     1441 tcctccccag cacttcctac acagcctgct gaagaagcag cacggaagag agaggtccgt
     1501 ctaatgaaga acagggaggc agcaagagaa tgtcgtagaa agaagaaaga atatgtgaaa
     1561 tgtttagaga acagagtggc agtgcttgaa aaccaaaaca aaacattgat tgaggagcta
     1621 aaagcactta aggaccttta ctgccacaaa tcagattaaa atcactagtg aattcgatat
     1681 caagcttatc gataccgtcg acctcgaggg ggggcccggt accttaatta attaaggtac
     1741 caggtaagtg tacccaattc gccctatagt gagtcgtatt acaattcact cgatcgccct
     1801 tcccaacagt tgcgcagcct gaatggcgaa tggagatcca atttttaagt gtataatgtg
     1861 ttaaactact gattctaatt gtttgtgtat tttagattca cagtcccaag gctcatttca
     1921 ggcccctcag tcctcacagt ctgttcatga tcataatcag ccataccaca tttgtagagg
     1981 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     2041 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2101 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2161 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     2221 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     2281 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     2341 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     2401 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     2461 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     2521 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     2581 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     2641 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2701 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2761 aggaagaatc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     2821 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2881 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2941 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3001 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3061 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3121 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     3181 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3241 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3301 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     3361 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     3421 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     3481 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     3541 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     3601 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagaacatca ggggctcgcg
     3661 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     3721 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     3781 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     3841 gatattgctg aagaacttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     3901 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     3961 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4021 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4081 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     4141 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     4201 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     4261 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     4321 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     4381 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     4441 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4501 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4561 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4621 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4681 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4741 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4801 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4861 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4921 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4981 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5041 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5101 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5161 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5221 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag
     5281 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccg
//
