LOCUS       pJM118-CFP-LacI-        7421 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 7421)
  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..7421
                     /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..4059
                     /codon_start=1
                     /label= CFP-LacI-MR-NTD
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKSGLRSMVKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTRE
                     KVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVV
                     VSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQT
                     PINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPI
                     AEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVG
                     YDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLA
                     PNTQTASPRALADSLMQLARQVSRSSLRPPKKKRKVGAPGSAGSAAGGSAIMETKGYHS
                     LPEGLDMERRWGQVSQAVERSSLGPTERTDENNYMEIVNVSCVSGAIPNNSTQGSSKEK
                     QELLPCLQQDNNRPGILTSDIKTELESKELSATVAESMGLYMDSVRDADYSYEQQNQQG
                     SMSPAKIYQNVEQLVKFYKGNGHRPSTLSCVNTPLRSFMSDSGSSVNGGVMRAVVKSPI
                     MCHEKSPSVCSPLNMTSSVCSPAGINSVSSTTASFGSFPVHSPITQGTPLTCSPNVENR
                     GSRSHSPAHASNVGSPLSSPLSSMKSSISSPPSHCSVKSPVSSPNNVTLRSSVSSPANI
                     NNSRCSVSSPSNTNNRSTLSSPAASTVGSICSPVNNAFSYTASGTSAGSSTLRDVVPSP
                     DTQEKGAQEVPFPKTEEVESAISNGVTGQLNIVQYIKPEPDGAFSSSCLGGNSKINSDS
                     SFSVPIKQESTKHSCSGTSFKGNPTVNPFPFMDGSYFSFMDDKDYYSLSGILGPPVPGF
                     DGNCEGSGFPVGIKQEPDDGSYYPEASIPSSAIVGVNSGGQSFHYRIGAQGTISLSRSA
                     RDQSFQHLSSFPPVNTLVESWKSHGDLSSRRSDGYPVLEYIPENVSSSTLRSVSTGSSR
                     PSKI"
     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..2277
                     /label=Fragment.FOR
     misc_feature    2248..2252
                     /label=MCS
     misc_feature    2254..4052
                     /label=MR_NTD
     primer_bind     complement(2915..2934)
                     /label=Mid_MR_NTD_RV
     primer_bind     2933..2952
                     /label=Mid_MR_NTD_FW
     primer_bind     3745..3764
                     /label=END_MR_NTD_FW
     primer_bind     complement(4023..4102)
                     /label=Fragment.REV
     misc_feature    4063..4075
                     /label=MCS
     primer_bind     complement(4150..4168)
                     /label=ValidateInsert_pJM118_RV
     polyA_signal    4209..4330
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4337..4792)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4819..4923
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4925..5282
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5133..5268
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5317..6111
                     /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    6343..6390
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6719..7307
                     /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 ataatggaga ccaaaggcta ccacagtctc
     2281 cctgaaggtc tagatatgga aagacggtgg ggtcaagttt ctcaggctgt ggagcgttct
     2341 tccctgggac ctacagagag gaccgatgag aataactaca tggagattgt caacgtaagc
     2401 tgtgtttccg gtgctattcc aaacaacagt actcaaggaa gcagcaaaga aaaacaagaa
     2461 ctactccctt gccttcagca agacaataat cggcctggga ttttaacatc tgatattaaa
     2521 actgagctgg aatctaagga actttcagca actgtagctg agtccatggg tttatatatg
     2581 gattctgtaa gagatgctga ctattcctat gagcagcaga accaacaagg aagcatgagt
     2641 ccagctaaga tttatcagaa tgttgaacag ctggtgaaat tttacaaagg aaatggccat
     2701 cgtccttcca ctctaagttg tgtgaacacg cccttgagat catttatgtc tgactctggg
     2761 agctccgtga atggtggcgt catgcgcgcc gttgttaaaa gccctatcat gtgtcatgag
     2821 aaaagcccgt ctgtttgcag ccctctgaac atgacatctt cggtttgcag ccctgctgga
     2881 atcaactctg tgtcctccac cacagccagc tttggcagtt ttccagtgca cagcccaatc
     2941 acccagggaa ctcctctgac atgctcccct aatgttgaaa atcgaggctc caggtcgcac
     3001 agccctgcac atgctagcaa tgtgggctct cctctctcaa gtccgttaag tagcatgaaa
     3061 tcctcaattt ccagccctcc aagtcactgc agtgtaaaat ctccagtctc cagtcccaat
     3121 aatgtcactc tgagatcctc tgtgtctagc cctgcaaata ttaacaactc aaggtgctct
     3181 gtttccagcc cttcgaacac taataacaga tccacgcttt ccagtccggc agccagtact
     3241 gtgggatcta tctgtagccc tgtaaacaat gccttcagct acactgcttc tggcacctct
     3301 gctggatcca gtacattgcg ggatgtggtt cccagtccag acacgcagga gaaaggtgct
     3361 caagaggtcc cttttcctaa gactgaggaa gtagagagtg ccatctcaaa tggtgtgact
     3421 ggccagctta atattgtcca gtacataaaa ccagaaccag atggagcttt tagcagctca
     3481 tgtctaggag gaaatagcaa aataaattcg gattcttcat tctcagtacc aataaagcaa
     3541 gaatcaacca agcattcatg ttcaggcacc tcttttaaag ggaatccaac agtaaacccg
     3601 tttccattta tggatggctc gtatttttcc tttatggatg ataaagacta ttattcccta
     3661 tcaggaattt taggaccacc tgtgcccggc tttgatggta actgtgaagg cagcggattc
     3721 ccagtgggta ttaaacaaga accagatgat gggagctatt acccagaggc cagcatccct
     3781 tcctctgcta ttgttggggt gaattcaggt ggacagtcct tccactacag gattggtgct
     3841 caaggtacaa tatctttatc acgatcggct agagaccaat ctttccaaca cctgagttcc
     3901 tttcctcctg tcaatacttt agtggagtca tggaaatcac acggcgacct gtcgtctaga
     3961 agaagtgatg ggtatccggt cttagaatac attccagaaa atgtatcaag ctctacttta
     4021 cgaagtgttt ctactggatc ttcaagacct tcaaaaatat gagtctagag ggcccgaatt
     4081 cgagcgccgt cgacaagctt gcggccgcaa tcagccatac cacatttgta gaggttttac
     4141 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     4201 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4261 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4321 atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     4381 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     4441 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     4501 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     4561 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     4621 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     4681 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     4741 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     4801 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4861 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4921 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4981 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     5041 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5101 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     5161 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     5221 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     5281 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5341 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5401 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5461 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     5521 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5581 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5641 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5701 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5761 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5821 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     5881 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5941 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     6001 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     6061 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     6121 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     6181 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     6241 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     6301 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     6361 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     6421 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     6481 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     6541 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     6601 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6661 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6721 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6781 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6841 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6901 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6961 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     7021 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     7081 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     7141 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     7201 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     7261 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     7321 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     7381 tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//