LOCUS       sequence-237488-        6037 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMVP destination vector, empty Sleeping Beauty transposon vector 
            backbone w/ eCFP selection cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6037)
  AUTHORS   Haldeman JM, Conway AE, Arlotto ME, Slentz DH, Muoio DM, Becker TC, 
            Newgard CB
  TITLE     Creation of versatile cloning platforms for transgene expression and
            dCas9-based epigenome editing.
  JOURNAL   Nucleic Acids Res. 2018 Dec 27. pii: 5264291. doi: 
            10.1093/nar/gky1286.
  PUBMED    30590691
REFERENCE   2  (bases 1 to 6037)
  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: "Nucleic 
            Acids Res. 2018 Dec 27. pii: 5264291. doi: 10.1093/nar/gky1286."
COMMENT     Sequence Label: KN302: pMVP/SB/eCFP-DEST
FEATURES             Location/Qualifiers
     source          1..6037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(129..197,198..989)
                     /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(347..366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1649..1668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1902..1919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1925..1942
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1926..1942
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2241..2365
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2390..2395,2396..2413,2414..2420)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2474..3133
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2540..2559)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3475..3780
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3659..3678
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3821..3945)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3960..3976)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3961..3977)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        3997..4313
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4164..4299
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4226..4245
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4361..4408
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(4427..4429,4430..4432,4433..5146)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4472..4493)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4733..4752)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5080..5101
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5186..5205)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5230..5278
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     complement(5494..5515)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5928..5947
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagctgtaaa acgacggcca gtagctccac cgcggtggcg gccgctctag aactagtgta
     1981 caccctatac agttgaagtc ggaagtttac atacacttaa gttggagtca ttaaaactcg
     2041 tttttcaact actccacaaa tttcttgtta acaaacaata gttttggcaa gtcagttagg
     2101 acatctactt tgtgcatgac acaagtcatt tttccaacaa ttgtttacag acagattatt
     2161 tcacttataa ttcactgtat cacaattcca gtgggtcaga agtttacata cactaagttg
     2221 ggctgcagga attcgatatc acaagtttgt acaaaaaagc tgaacgagaa acgtaaaatg
     2281 atataaatat caatatatta aattagattt tgcataaaaa acagactaca taatactgta
     2341 aaacacaaca tatccagtca ctatggcggc cgcattaggc accccaggct ttacacttta
     2401 tgcttccggc tcgtataatg tgtggatttt gagttaggat ccgtcgagat tttcaggagc
     2461 taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg
     2521 gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac
     2581 cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta
     2641 tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat tccgtatggc
     2701 aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca
     2761 tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt
     2821 tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa
     2881 agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt
     2941 tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata
     3001 ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg
     3061 tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca
     3121 gggcggggcg taaacgcgtg gatccggctt actaaaagcc agataacagt atgcgtattt
     3181 gcgcgctgat ttttgcggta taagaatata tactgatatg tatacccgaa gtatgtcaaa
     3241 aagaggtatg ctatgaagca gcgtattaca gtgacagttg acagcgacag ctatcagttg
     3301 ctcaaggcat atatgatgtc aatatctccg gtctggtaag cacaaccatg cagaatgaag
     3361 cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg gctgaggtcg
     3421 cccggtttat tgaaatgaac ggctcttttg ctgacgagaa caggggctgg tgaaatgcag
     3481 tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt
     3541 gatattattg acacgcccgg gcgacggatg gtgatccccc tggccagtgc acgtctgctg
     3601 tcagataaag tctcccgtga actttacccg gtggtgcata tcggggatga aagctggcgc
     3661 atgatgacca ccgatatggc cagtgtgccg gtctccgtta tcggggaaga agtggctgat
     3721 ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa
     3781 atgtcaggct cccttataca cagccagtct gcaggtcgac catagtgact ggatatgttg
     3841 tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata tattgatatt
     3901 tatatcattt tacgtttctc gttcagcttt cttgtacaaa gtggtgatat caagcttatc
     3961 gataccgtcg acctcgacat ggtcatagct gttgacggtg tggaaagtcc ccaggctccc
     4021 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4081 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4141 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4201 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4261 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4321 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
     4381 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg tgagcaaggg
     4441 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     4501 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     4561 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     4621 gacctggggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     4681 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     4741 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     4801 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     4861 ctacatcagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa
     4921 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     4981 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     5041 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     5101 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaacgta ccggttagta
     5161 atgagtttaa acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     5221 gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat
     5281 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     5341 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     5401 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatcc ggttaaatca
     5461 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactgtac
     5521 attgagtgta tgtaaacttc tgacccactg ggaatgtgat gaaagaaata aaagctgaaa
     5581 tgaatcattc tctctactat tattctgata tttcacattc ttaaaataaa gtggtgatcc
     5641 taactgacct aagacaggga atttttacta ggattaaatg tcaggaattg tgaaaaagtg
     5701 agtttaaatg tatttggcta aggtgtatgt aaacttccga cttcaactgt atagggattg
     5761 tacaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     5821 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     5881 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     5941 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     6001 cccgccgcgc ttaatgcgcc gctacagggc gcgtcag
//
