LOCUS       pCH301_LCGC14_In        5458 bp DNA     circular SYN 12-AUG-2025
DEFINITION  Ligation of pCH298* into pCH83*.
ACCESSION   .
VERSION     .
KEYWORDS    pCH301
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5458)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 12, 2025 from SnapGene Viewer 6.1.2
            https://www.snapgene.com
COMMENT     Vector molecule:  pCH83*
               Fragment ends: KpnI and NdeI
               Fragment size: 5317
            Insert molecule:  pCH298*
               Fragment ends: NdeI and KpnI
               Fragment size: 141
FEATURES             Location/Qualifiers
     source          1..5458
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..57)
                     /label=pTrcHis-RP
                     /note="color: black; sequence: CTGATTTAATCTGTATCAGG"
     primer_bind     complement(130..149)
                     /label=pTI2-1-2
                     /note="color: black; sequence: GCGTTTCACTTCTGAGTTCG"
     primer_bind     complement(131..150)
                     /label=pKK223-3-for
                     /note="color: black; sequence: GGCGTTTCACTTCTGAGTTC"
     CDS             522..1382
                     /codon_start=1
                     /label=Bla
                     /note="color: #cc0000"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(567..586)
                     /label=AmpStart
                     /note="color: black; sequence: AAAACAGGAAGGCAAAATGC"
     primer_bind     1313..1332
                     /label=AmpStop
                     /note="color: black; sequence: TCAGGCAACTATGGATGAAC"
     primer_bind     2965..2984
                     /label=GEX3-rev
                     /note="color: black; sequence: GCTTACAGACAAGCTGTGAC"
     primer_bind     2993..3011
                     /label=pGEX-3'
                     /note="color: black; sequence: GGAGCTGCATGTGTCAGAG"
     primer_bind     2994..3012
                     /label=GEX3-rev-25
                     /note="color: black; sequence: GAGCTGCATGTGTCAGAGG"
     primer_bind     3143..3159
                     /label=pBR3
                     /note="color: black; sequence: TCCCCATCGGTGATGTC"
     primer_bind     3212..3226
                     /label=pET-up
                     /note="color: black; sequence: ATGCGTCCGGCGTAG"
     CDS             complement(3353..4282)
                     /codon_start=1
                     /label=araC
                     /note="color: #cc0000"
                     /translation="MQYGQLVSSLNGGSMKSMAEAQNDPLLPGYSFNAHLVAGLTPIEA
                     NGYLDFFIDRPLGMKGYILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPE
                     AREWYHQWVYFRPRAYWHEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGR
                     YSELLAINLLEQLLLRRMEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVC
                     LSPSRLSHLFRQQLGISVLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRV
                     FKKCTGASPSEFRAGCEEKVNDVAVKLS"
     misc_feature    4382..4410
                     /label=araC Promoter
                     /note="color: #cc0000; direction: RIGHT"
     primer_bind     4465..4484
                     /label=pBAD-for
                     /note="color: black; sequence: ATGCCATAGCATTTTTATCC"
     primer_bind     4465..4484
                     /label=pBAD-FP
                     /note="color: black; sequence: ATGCCATAGCATTTTTATCC"
     misc_feature    4507..4534
                     /label=pBAD Promoter
                     /note="color: #cc0000; direction: RIGHT"
     primer_bind     4515..4534
                     /label=pBAD-F+50
                     /note="color: black; sequence: TTATCGCAACTCTCTACTGT"
     CDS             4599..5456
                     /codon_start=1
                     /label=pCH301
                     /note="color: #cc0000"
                     /translation="MVEKLKIQKIEKLEDFDNEYVYDISVDKETPYFFGNNILVHNSAY
                     GTSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTTGKLPVPWP
                     TLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGTYKTRAEVKFEGD
                     TLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKANFKIRHNVEDGSV
                     QLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEFVTAAGITHGGSH
                     HHHHH"
     CDS             4599..4724
                     /codon_start=1
                     /label=LCGC14 IntC
                     /note="color: #cc0000"
                     /translation="MVEKLKIQKIEKLEDFDNEYVYDISVDKETPYFFGNNILVHN"
     misc_feature    4725..4733
                     /label=ExtC
                     /note="color: #cc0000; direction: RIGHT"
     misc_feature    4740..5432
                     /label=sfGFP
                     /note="color: #cc0000; direction: RIGHT"
     primer_bind     complement(5415..5455)
                     /label=oCH150
                     /note="IntC-sfGFP-His6 (rev)"
                     /note="color: black; sequence: 
                     TGATGGTGATGGTGATGGCTGCCGCCATGAGTGATACCCGC"
     CDS             5439..5456
                     /codon_start=1
                     /label=His6
                     /note="color: #cc0000"
                     /translation="HHHHHH"
ORIGIN
        1 agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac
       61 gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg
      121 accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtccccca
      181 tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtgc aaagactggg
      241 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
      301 gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
      361 aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
      421 tacaaactct ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
      481 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
      541 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
      601 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
      661 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
      721 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
      781 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
      841 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
      901 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
      961 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     1021 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     1081 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     1141 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     1201 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     1261 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     1321 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     1381 aactgtcaga ccaagtttac tcatatatac tttagattga tttacgcgcc ctgtagcggc
     1441 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1501 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1561 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     1621 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     1681 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     1741 ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     1801 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     1861 atattaacgt ttacaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1921 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1981 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2041 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2101 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     2161 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2221 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2281 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2341 gcgaacgacc tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct
     2401 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2461 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2521 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2581 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2641 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2701 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2761 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc ggatatggtg
     2821 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     2881 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     2941 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3001 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagcaagg agatggcgcc
     3061 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     3121 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     3181 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga tctaattctc
     3241 atgtttgaca gcttatcatc gatgcataat gtgcctgtca aatggacgaa gcagggattc
     3301 tgcaaaccct atgctactcc gtcaagccgt caattgtctg attcgttacc aattatgaca
     3361 acttgacggc tacatcattc actttttctt cacaaccggc acggaactcg ctcgggctgg
     3421 ccccggtgca ttttttaaat acccgcgaga aatagagttg atcgtcaaaa ccaacattgc
     3481 gaccgacggt ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc tggctgatac
     3541 gttggtcctc gcgccagctt aagacgctaa tccctaactg ctggcggaaa agatgtgaca
     3601 gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg
     3661 ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg attatccatc ggtggatgga
     3721 gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga tttatcgcca
     3781 gcagctccga atagcgccct tccccttgcc cggcgttaat gatttgccca aacaggtcgc
     3841 tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc cgtattggca aatattgacg
     3901 gccagttaag ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc
     3961 attcgcgagc ctccggatga cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa
     4021 tatcacccgg tcggcaaaca aattctcgtc cctgattttt caccaccccc tgaccgcgaa
     4081 tggtgagatt gagaatataa cctttcattc ccagcggtcg gtcgataaaa aaatcgagat
     4141 aaccgttggc ctcaatcggc gttaaacccg ccaccagatg ggcattaaac gagtatcccg
     4201 gcagcagggg atcattttgc gcttcagcca tacttttcat actcccgcca ttcagagaag
     4261 aaaccaattg tccatattgc atcagacatt gccgtcactg cgtcttttac tggctcttct
     4321 cgctaaccaa accggtaacc ccgcttatta aaagcattct gtaacaaagc gggaccaaag
     4381 ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg cagaaaagtc cacattgatt
     4441 atttgcacgg cgtcacactt tgctatgcca tagcattttt atccataaga ttagcggatc
     4501 ctacctgacg ctttttatcg caactctcta ctgtttctcc atacccgttt ttttgggcta
     4561 gaaataattt tgtttaactt taagaaggag atatacatat ggtggagaaa ttgaagattc
     4621 aaaagattga gaaactggaa gactttgaca acgagtatgt ctatgacatc tccgtggata
     4681 aagagacgcc atatttcttc ggcaataaca tccttgttca taacagcgct tatggtacca
     4741 gcaaaggcga agaactgttt accggcgtgg tgccgattct ggttgaactg gatggcgatg
     4801 ttaatggcca taagtttagc gtgcgcggcg aaggcgaagg cgatgcgacc aacggcaaac
     4861 tgaccctgaa gtttatttgc accaccggca aactgccggt gccgtggccg accctggtga
     4921 ccaccctgac ctatggcgtg cagtgcttta gccgctatcc ggatcacatg aaacgccatg
     4981 atttctttaa gtccgcgatg ccggaaggct atgttcagga acgcaccatt agctttaaag
     5041 atgatggcac ctataagacc cgcgcggaag tgaaatttga aggcgatacc ctggtgaacc
     5101 gcattgaact gaaaggcatt gacttcaaag aagatggcaa cattctgggc cataaactgg
     5161 agtacaattt caacagccat aacgtgtata ttaccgcgga caaacagaag aacggcatca
     5221 aagcgaattt caaaatccgc cataatgtgg aagatggcag cgtgcagctg gcggatcact
     5281 atcagcagaa taccccgatc ggcgatggcc cggtgctgct gccggacaat cactacctga
     5341 gcacccagtc cgtgctgagc aaagatccga atgagaaacg cgatcacatg gttctgctgg
     5401 agtttgtgac cgcggcgggt atcactcatg gcggcagcca tcaccatcac catcacta
//
