LOCUS       sequence-482901-        4693 bp    DNA     circular SYN 12-AUG-2025
DEFINITION  Localization: C2 Cloning Vector, Excitation: 508 / 572, Emission: 
            518 / 580.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4693)
  AUTHORS   Davidson Lab
  TITLE     Michael Davidson Empty Backbones
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 4693)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 12, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "unpublished"
COMMENT     Sequence Label: Kaede-C2
FEATURES             Location/Qualifiers
     source          1..4693
                     /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"
     CDS             553..1227
                     /codon_start=1
                     /product="Kaede green-to-red photoconvertible fluorescent 
                     protein (Ando et al., 2002)"
                     /label=Kaede
                     /translation="MSLIKPEMKIKLLMEGNVNGHQFVIEGDGKGHPFEGKQSMDLVVK
                     EGAPLPFAYDILTTAFHYGNRVFAKYPDHIPDYFKQSFPKGFSWERSLMFEDGGVCIAT
                     NDITLKGDTFFNKVRFDGVNFPPNGPVMQKKTLKWEASTEKMYLRDGVLTGDITMALLL
                     KGDVHYRCDFRTTYKSRQEGVKLPGYHFVDHCISILRHDKDYNEVKLYEHAVAHSGLPD
                     NVK"
     misc_feature    1241..1297
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    1421..1542
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1458..1477)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1512..1531
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1549..2004)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1686..1707)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1898..1917
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2031..2135
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2137..2494
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2137..2328
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2160..2180)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2345..2480
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2529..3323
                     /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             2529..3323
                     /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(2583..2602)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3193..3212
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3511..3530)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3555..3602
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3931..4519
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4420..4439
                     /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 gctagcgcta
      541 ccggtcgcca ccatgagtct gattaaacca gaaatgaaga tcaagctgct tatggaaggc
      601 aatgtaaacg ggcaccagtt tgttattgag ggagatggaa aaggccatcc ttttgaggga
      661 aaacagagta tggaccttgt agtcaaagaa ggcgcacctc tcccttttgc ctacgatatc
      721 ttgacaacag cattccatta tggtaacagg gtttttgcta aatacccaga ccatatacca
      781 gactacttca agcagtcgtt tcccaaaggg ttttcttggg agcgaagcct gatgttcgag
      841 gacgggggcg tttgcatcgc tacaaatgac ataacactga aaggagacac tttttttaac
      901 aaagttcgat ttgatggcgt aaactttccc ccaaatggtc ctgttatgca gaagaagact
      961 ctgaaatggg aggcatccac tgagaaaatg tatttgcgtg atggagtgtt gacgggcgat
     1021 attacaatgg ctctgctgct taaaggagat gtccattacc gatgtgactt cagaactact
     1081 tacaaatcta ggcaggaggg tgtcaagttg ccaggatatc actttgtcga tcactgcatc
     1141 agcatattga ggcatgacaa agactacaac gaggttaagc tgtatgagca cgctgttgcc
     1201 cattctggat tgccggacaa cgtcaagtcc ggccggactc agatctcgag ctcaagcttc
     1261 gaattctgca gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat
     1321 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     1381 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     1441 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     1501 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
     1561 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     1621 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     1681 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     1741 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     1801 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     1861 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     1921 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     1981 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2041 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2101 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     2161 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2221 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2281 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     2341 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     2401 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     2461 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     2521 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     2581 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     2641 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     2701 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     2761 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     2821 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     2881 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     2941 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3001 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     3061 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3121 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3181 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     3241 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     3301 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     3361 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     3421 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     3481 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     3541 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     3601 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     3661 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     3721 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     3781 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3841 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3901 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3961 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4021 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4081 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4141 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4201 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4261 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4321 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4381 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4441 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4501 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4561 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4621 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     4681 atatggagtt ccg
//
