LOCUS       sequence-393320-        4731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: C1 Cloning Vector, Excitation: 400 / 504, Emission: 
            515 / 517.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4731)
  AUTHORS   Davidson Lab
  TITLE     Michael Davidson Empty Backbones
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 4731)
  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            "unpublished"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: mPA-GFP-C1
FEATURES             Location/Qualifiers
     source          1..4731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..32)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     623..642
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(941..960)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    985..1032
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1361..1949
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1850..1869
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2124..2427
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2428..2631
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2582..2602
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2670..2679
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2676..2678,2679..2681,2682..3392)
                     /codon_start=1
                     /product="photoactivatable GFP variant (Patterson et al., 
                     2002)"
                     /label=PA-GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2721..2742)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2982..3001)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3329..3350
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3393..3458
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3582..3703
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3619..3638)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3673..3692
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3710..4165)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3847..3868)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4059..4078
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4192..4296
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4298..4655
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4298..4489
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4321..4341)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4506..4641
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(4690..4731,1..753)
                     /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             join(4690..4731,1..753)
                     /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"
ORIGIN
        1 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
       61 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
      121 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
      181 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
      241 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
      301 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
      361 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
      421 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
      481 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
      541 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
      601 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
      661 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
      721 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
      781 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
      841 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
      901 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
      961 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     1021 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     1081 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     1141 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     1201 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     1261 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1321 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1381 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1441 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1501 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     1561 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     1621 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     1681 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     1741 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     1801 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     1861 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     1921 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     1981 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2041 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     2101 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     2161 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     2221 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     2281 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     2341 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     2401 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     2461 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     2521 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     2581 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
     2641 aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt
     2701 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     2761 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     2821 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc agctacggcg
     2881 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2941 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     3001 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     3061 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     3121 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
     3181 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     3241 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagccaccag tccaagctga
     3301 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     3361 ggatcactct cggcatggac gagctgtaca agtccggact cagatctcga gctcaagctt
     3421 cgaattctgc agtcgacggt accgcgggcc cgggatccac cggatctaga taactgatca
     3481 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     3541 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     3601 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3661 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc
     3721 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3781 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3841 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3901 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3961 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4021 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4081 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4141 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4201 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4261 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     4321 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4381 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4441 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4501 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4561 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     4621 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     4681 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc t
//
