LOCUS       pJM118-CFP-LacI-        7316 bp DNA     circular SYN 20-OCT-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7316)
  AUTHORS   Software Licensing Group
  TITLE     Direct Submission
  JOURNAL   Exported Oct 20, 2025 from SnapGene 7.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        19..348
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     regulatory      367..372
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             373..3951
                     /codon_start=1
                     /label= CFP-LacI-PR-NTD
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKSGLRSMVKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTRE
                     KVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVV
                     VSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQT
                     PINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPI
                     AEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVG
                     YDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLA
                     PNTQTASPRALADSLMQLARQVSRSSLRPPKKKRKVGAPGSAGSAAGGSAIMTELKAKG
                     PRAPHVAGGPPSPEVGSPLLCRPAAGPFPGSQTSDTLPEVSAIPISLDGLLFPRPCQGQ
                     DPSDEKTQDQQSLSDVEGAYSRAEATRGAGGSSSSPPEKDSGLLDSVLDTLLAPSGPGQ
                     SQPSPPACEVTSSWCLFGPELPEDPPAAPATQRVLSPLMSRSGCKVGDSSGTAAAHKVL
                     PRGLSPARQLLLPASESPHWSGAPVKPSPQAAAVEVEEEDGSESEESAGPLLKGKPRAL
                     GGAAAGGGAAAVPPGAAAGGVALVPKEDSRFSAPRVALVEQDAPMAPGRSPLATTVMDF
                     IHVPILPLNHALLAARTRQLLEDESYDGGAGAASAFAPPRTSPCASSTPVAVGDFPDCA
                     YPPDAEPKDDAYPLYSDFQPPALKIKEEEEGAEASARSPRSYLVAGANPAAFPDFPLGP
                     PPPLPPRATPSRPGEAAVTAAPASASVSSASSSGSTLECILYKAEGAPPQQGPFAPPPC
                     KAPGASGCLLPRDGLPSTSASAAAAGAAPALYPALGLNGLPQLGYQAAVLKEGLPQVYP
                     PYLNYLRPDSEASQSPQYSFESLPQKI"
     CDS             373..1089
                     /codon_start=1
                     /label=ECFP coding
                     /label=ECFP\coding
                     /note="vntifkey=21"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    1075..1089
                     /label=right flanker
     CDS             1105..2208
                     /codon_start=1
                     /label=LacI
                     /translation="MVKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE
                     LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSG
                     VEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFS
                     HEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWS
                     AMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSS
                     CYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASP
                     RALADSLMQLARQVSRSSLRPPKKKRKV"
     primer_bind     2134..2153
                     /label=Validate insert pJM118_FW
     CDS             2188..2208
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     2213..2275
                     /label=Fragment.FOR
     misc_feature    2248..2252
                     /label=MCS
     misc_feature    2254..3951
                     /label=PR_NTD
     primer_bind     2920..2939
                     /label=PR_NTD_1_FW
     primer_bind     complement(2920..2939)
                     /label=PR_NTD_1_RV
     primer_bind     complement(3924..3997)
                     /label=Fragment.REV
     misc_feature    3955..3967
                     /label=MCS
     primer_bind     complement(4045..4063)
                     /label=ValidateInsert_pJM118_RV
     polyA_signal    4104..4225
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4232..4687)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4714..4818
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4820..5177
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5028..5163
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5212..6006
                     /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"
     polyA_signal    6238..6285
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6614..7202
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tagttattac tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
       61 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
      121 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
      181 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
      241 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
      301 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctagcgcta
      361 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      421 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      481 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      541 ccctggccca ccctcgtgac caccctgacc tggggcgtgc agtgcttcag ccgctacccc
      601 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      661 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      721 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      781 atcctggggc acaagctgga gtacaactac atcagccaca acgtctatat caccgccgac
      841 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcagc
      901 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
      961 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1021 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1081 ctgtacaagt ccggactcag atccatggtg aaaccagtaa cgttatacga tgtcgcagag
     1141 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
     1201 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
     1261 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
     1321 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
     1381 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
     1441 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
     1501 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
     1561 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg
     1621 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
     1681 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
     1741 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
     1801 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
     1861 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
     1921 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag
     1981 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
     2041 gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg
     2101 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     2161 caggtttccc gaagcagcct gaggcctcct aagaagaaga ggaaggttgg tgcaccgggt
     2221 tctgcgggtt ctgccgcagg tggatccgcg ataatgactg agctgaaggc aaagggtccc
     2281 cgggctcccc acgtggcggg cggcccgccc tcccccgagg tcggatcccc actgctgtgt
     2341 cgcccagccg caggtccgtt cccggggagc cagacctcgg acaccttgcc tgaagtttcg
     2401 gccataccta tctccctgga cgggctactc ttccctcggc cctgccaggg acaggacccc
     2461 tccgacgaaa agacgcagga ccagcagtcg ctgtcggacg tggagggcgc atattccaga
     2521 gctgaagcta caaggggtgc tggaggcagc agttctagtc ccccagaaaa ggacagcgga
     2581 ctgctggaca gtgtcttgga cactctgttg gcgccctcag gtcccgggca gagccaaccc
     2641 agccctcccg cctgcgaggt caccagctct tggtgcctgt ttggccccga acttcccgaa
     2701 gatccaccgg ctgcccccgc cacccagcgg gtgttgtccc cgctcatgag ccggtccggg
     2761 tgcaaggttg gagacagctc cgggacggca gctgcccata aagtgctgcc ccggggcctg
     2821 tcaccagccc ggcagctgct gctcccggcc tctgagagcc ctcactggtc cggggcccca
     2881 gtgaagccgt ctccgcaggc cgctgcggtg gaggttgagg aggaggatgg ctctgagtcc
     2941 gaggagtctg cgggtccgct tctgaagggc aaacctcggg ctctgggtgg cgcggcggct
     3001 ggaggaggag ccgcggctgt cccgccgggg gcggcagcag gaggcgtcgc cctggtcccc
     3061 aaggaagatt cccgcttctc agcgcccagg gtcgccctgg tggagcagga cgcgccgatg
     3121 gcgcccgggc gctccccgct ggccaccacg gtgatggatt tcatccacgt gcctatcctg
     3181 cctctcaatc acgccttatt ggcagcccgc actcggcagc tgctggaaga cgaaagttac
     3241 gacggcgggg ccggggctgc cagcgccttt gccccgccgc ggacttcacc ctgtgcctcg
     3301 tccaccccgg tcgctgtagg cgacttcccc gactgcgcgt acccgcccga cgccgagccc
     3361 aaggacgacg cgtaccctct ctatagcgac ttccagccgc ccgctctaaa gataaaggag
     3421 gaggaggaag gcgcggaggc ctccgcgcgc tccccgcgtt cctaccttgt ggccggtgcc
     3481 aaccccgcag ccttcccgga tttcccgttg gggccaccgc ccccgctgcc gccgcgagcg
     3541 accccatcca gacccgggga agcggcggtg acggccgcac ccgccagtgc ctcagtctcg
     3601 tctgcgtcct cctcggggtc gaccctggag tgcatcctgt acaaagcgga gggcgcgccg
     3661 ccccagcagg gcccgttcgc gccgccgccc tgcaaggcgc cgggcgcgag cggctgcctg
     3721 ctcccgcggg acggcctgcc ctccacctcc gcctctgccg ccgccgccgg ggcggccccc
     3781 gcgctctacc ctgcactcgg cctcaacggg ctcccgcagc tcggctacca ggccgccgtg
     3841 ctcaaggagg gcctgccgca ggtctacccg ccctatctca actacctgag gccggattca
     3901 gaagccagcc agagcccaca atacagcttc gagtcattac ctcagaagat ttgagtctag
     3961 agggccctag gaattcgagc gccgtcgaca agcttgcggc cgcaatcagc cataccacat
     4021 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     4081 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     4141 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4201 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     4261 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     4321 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     4381 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     4441 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     4501 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     4561 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     4621 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     4681 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4741 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4801 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4861 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4921 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4981 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     5041 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5101 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5161 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     5221 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     5281 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     5341 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     5401 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5461 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5521 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5581 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5641 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5701 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     5761 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5821 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5881 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5941 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     6001 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     6061 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     6121 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     6181 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     6241 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     6301 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     6361 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     6421 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     6481 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6541 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6601 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6661 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6721 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6781 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6841 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6901 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6961 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     7021 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     7081 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7141 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     7201 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7261 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//