LOCUS       pCH308_LCGC14_MB        7014 bp DNA     circular SYN 12-AUG-2025
DEFINITION  C24S in pCH300.
ACCESSION   .
VERSION     .
KEYWORDS    pCH308
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7014)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 12, 2025 from SnapGene Viewer 6.1.2
            https://www.snapgene.com
COMMENT     Vector molecule:  pTT04*
               Fragment ends: BamHI and EcoRI
               Fragment size: 6639
            Insert molecule:  pCH298*
               Fragment ends: EcoRI and BamHI
               Fragment size: 375
FEATURES             Location/Qualifiers
     source          1..7014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             81..1163
                     /codon_start=1
                     /label=lacI
                     /note="color: #cc0000"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1185..1201
                     /label=PCR2
                     /note="color: black; sequence: TTAGCTCACTCATTAGG"
     primer_bind     1376..1395
                     /label=pKK223-3-rev
                     /note="color: black; sequence: CGGTTCTGGCAAATATTCTG"
     primer_bind     1383..1403
                     /label=pTI2-1-1
                     /note="color: black; sequence: GGCAAATATTCTGAAATGAGC"
     CDS             1406..1433
                     /codon_start=1
                     /label=Ptac promotor
                     /note="color: #cc0000"
                     /translation="LTINHRLV"
     primer_bind     1423..1444
                     /label=BAC1
                     /note="color: black; sequence: GCTCGTATGTTGTGTGGAATTG"
     primer_bind     1438..1456
                     /label=pET-T7side
                     /note="color: black; sequence: GGGAATTGTGAGCGGATAAC"
     primer_bind     1449..1468
                     /label=LacOp-for
                     /note="color: black; sequence: CGGATAACAATTTCACACAG"
     primer_bind     1449..1468
                     /label=pQE-FP
                     /note="color: black; sequence: CGGATAACAATTTCACACAG"
     CDS             1528..3096
                     /codon_start=1
                     /label=pCH300
                     /note="color: #cc0000"
                     /translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAIEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQTNSSSNNNNNNNNNNLGIEGRISEFDTDSVDAD
                     TIIKTNYGEMTIENLFKSSSIKGPSWAIDDQEFTIYDQIQILTYDPKTNEEIYRPFEYV
                     YRHKVSKPRWKIIDENGNEIILTNDHSVMIERDGKLIEAKPSEINPDTDILITIGEGSH
                     HHHHH"
     CDS             1528..2695
                     /codon_start=1
                     /label=orf'
                     /note="color: #cc0000"
                     /translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAIEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQTNSSSNNNNNNNNNNLGIEGRIS"
     CDS             1528..2627
                     /codon_start=1
                     /label=MBP
                     /note="color: #cc0000"
                     /translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAIEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQ"
     misc_feature    1858..1860
                     /label=V->I
                     /note="color: #cc0000; direction: RIGHT"
     primer_bind     2587..2605
                     /label=malE
                     /note="color: black; sequence: GGTCGTCAGACTGTCGATG"
     primer_bind     2592..2609
                     /label=pMalE
                     /note="color: black; sequence: TCAGACTGTCGATGAAGC"
     misc_feature    2677..2688
                     /label=Factor Xa Site
                     /note="color: #cc0000; direction: RIGHT"
     misc_feature    2701..2709
                     /label=ExtN
                     /note="color: #cc0000; direction: RIGHT"
     misc_feature    2710..3069
                     /label=LCGC14 IntN
                     /note="color: #cc0000; direction: RIGHT"
     primer_bind     2751..2799
                     /label=oCH457
                     /note="MBP-LCGC14N(C24S)-His6 (fwd)"
                     /note="color: black; sequence: 
                     GATGACTATCGAAAATTTGTTTAAGTCTAGTAGCATTAAAGGTCCGTCC"
     primer_bind     complement(2751..2799)
                     /label=oCH458
                     /note="MBP-LCGC14N(C24S)-His6 (rev)"
                     /note="color: black; sequence: 
                     GGACGGACCTTTAATGCTACTAGACTTAAACAAATTTTCGATAGTCATC"
     misc_feature    2779..2781
                     /label=C24S
                     /note="color: #cc0000; direction: RIGHT"
     CDS             3076..3093
                     /codon_start=1
                     /label=His6
                     /note="color: #cc0000"
                     /translation="HHHHHH"
     primer_bind     complement(3104..3121)
                     /label=M13-FP
                     /note="color: black; sequence: TGTAAAACGACGGCCAGT"
     primer_bind     complement(3105..3121)
                     /label=M13
                     /note="color: black; sequence: TGTAAAACGACGGCCAG"
     primer_bind     complement(3107..3124)
                     /label=UP-2
                     /note="color: black; sequence: CGTTGTAAAACGACGGCC"
     primer_bind     complement(3114..3135)
                     /label=BAC2
                     /note="color: black; sequence: CCCAGTCACGACGTTGTAAAAC"
     primer_bind     complement(3124..3140)
                     /label=M13-40
                     /note="color: black; sequence: GTTTTCCCAGTCACGAC"
     primer_bind     complement(3153..3172)
                     /label=pUCM13-52
                     /note="color: black; sequence: GCTGCAAGGCGATTAAGTTG"
     primer_bind     complement(3164..3180)
                     /label=EGF2
                     /note="color: black; sequence: GGGGATGTGCTGCAAGG"
     primer_bind     complement(3198..3214)
                     /label=PCR1
                     /note="color: black; sequence: CGGGCCTCTTCGCTATT"
     primer_bind     complement(3198..3213)
                     /label=PCR1minus1
                     /note="color: black; sequence: GGGCCTCTTCGCTATT"
     primer_bind     complement(3299..3318)
                     /label=pTrcHis-RP
                     /note="color: black; sequence: CTGATTTAATCTGTATCAGG"
     primer_bind     complement(3391..3410)
                     /label=pTI2-1-2
                     /note="color: black; sequence: GCGTTTCACTTCTGAGTTCG"
     primer_bind     complement(3392..3411)
                     /label=pKK223-3-for
                     /note="color: black; sequence: GGCGTTTCACTTCTGAGTTC"
     CDS             3786..4646
                     /codon_start=1
                     /label=bla
                     /note="color: #cc0000"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3831..3850)
                     /label=AmpStart
                     /note="color: black; sequence: AAAACAGGAAGGCAAAATGC"
     primer_bind     4577..4596
                     /label=AmpStop
                     /note="color: black; sequence: TCAGGCAACTATGGATGAAC"
     primer_bind     6289..6308
                     /label=GEX3-rev
                     /note="color: black; sequence: GCTTACAGACAAGCTGTGAC"
     primer_bind     6317..6335
                     /label=pGEX-3'
                     /note="color: black; sequence: GGAGCTGCATGTGTCAGAG"
     primer_bind     6318..6336
                     /label=GEX3-rev-25
                     /note="color: black; sequence: GAGCTGCATGTGTCAGAGG"
ORIGIN
        1 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
       61 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
      121 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
      181 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
      241 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
      301 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
      361 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
      421 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
      481 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
      541 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
      601 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
      661 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
      721 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
      781 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
      841 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
      901 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
      961 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     1021 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     1081 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     1141 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
     1201 gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg
     1261 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
     1321 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
     1381 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga
     1441 attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt gttttcacga
     1501 gcacttcacc aacaaggacc atagcatatg aaaactgaag aaggtaaact ggtaatctgg
     1561 attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat
     1621 accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt
     1681 gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac
     1741 gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat
     1801 ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctatt
     1861 gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa
     1921 gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac
     1981 ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag
     2041 tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg
     2101 ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac
     2161 tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg
     2221 gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc
     2281 aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt
     2341 ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg
     2401 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag
     2461 ttggcgaaag atccacgtat tgccgccacc atggaaaacg cccagaaagg tgaaatcatg
     2521 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc
     2581 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg
     2641 aacaacaaca acaataacaa taacaacaac ctcgggatcg agggaaggat ttcagaattc
     2701 gataccgata gcgtggacgc tgatacaatt attaagacga actacggaga gatgactatc
     2761 gaaaatttgt ttaagtctag tagcattaaa ggtccgtcct gggcaatcga cgatcaggaa
     2821 tttaccattt atgaccagat ccaaattttg acttacgacc caaagacgaa cgaggaaatt
     2881 taccgtccct tcgaatatgt atatcgtcat aaggtctcta agcctcgctg gaaaatcatc
     2941 gacgaaaatg ggaacgagat tattttaacg aacgaccact ccgttatgat tgagcgcgat
     3001 ggtaaactta ttgaggcgaa gccgagcgaa atcaacccgg acacggatat tctgattact
     3061 attggtgagg gatcccatca tcatcaccat cattaagctt ggcactggcc gtcgttttac
     3121 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     3181 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     3241 gcagcctgaa tggcgaatgg cagcttggct gttttggcgg atgagataag attttcagcc
     3301 tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca
     3361 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg
     3421 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
     3481 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     3541 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
     3601 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
     3661 acggatggcc tttttgcgtt tctacaaact ctttttgttt atttttctaa atacattcaa
     3721 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3781 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3841 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3901 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3961 gccccgaaga acgttctcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4021 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4081 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4141 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4201 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4261 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4321 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4381 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4441 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4501 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4561 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4621 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4681 ttgatttacc ccggttgata atcagaaaag ccccaaaaac aggaagattg tataagcaaa
     4741 tatttaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     4801 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     4861 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     4921 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     4981 acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     5041 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     5101 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     5161 caccacaccc gccgcgctta atgcgccgct acagggcgcg taaaaggatc taggtgaaga
     5221 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5281 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5341 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5401 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     5461 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5521 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5581 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5641 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5701 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5761 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5821 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5881 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5941 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6001 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     6061 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     6121 gtatttcaca ccgcatatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     6181 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     6241 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     6301 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     6361 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgca gcgattcaca gatgtctgcc
     6421 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
     6481 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
     6541 ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
     6601 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
     6661 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
     6721 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
     6781 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
     6841 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
     6901 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
     6961 gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga aatt
//
