LOCUS       Exported                5409 bp DNA     circular SYN 10-JUL-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pJCX4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5409)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 10, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      109..697
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     enhancer        872..1175
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1176..1379
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             1450..2043
                     /codon_start=1
                     /gene="Mycobacterium xenopi gyrA"
                     /product="modified GyrA intein (Southworth et al., 1999)"
                     /label=Mxe GyrA intein
                     /note="N-terminal cleavage of this intein can be induced by
                     thiols."
                     /translation="CITGDALVALPEGESVRIADIVPGARPNSDNAIDLKVLDRHGNPV
                     LADRLFHSGEHPVYTVRTVEGLRVTGTANHPLLCLVDVAGVPTLLWKLIDEIKPGDYAV
                     IQRSAFSVDCAGFARGKPEFAPTTYTVGVPGLVRFLEAHHRDPDAQAIADELTDGRFYY
                     AKVASVTDAGVQPVYSLRVDTADHAFITNGFVSHA"
     CDS             2074..2229
                     /codon_start=1
                     /gene="Bacillus circulans chiA"
                     /product="chitin binding domain from chitinase A1 (Watanabe
                     et al., 1994)"
                     /label=CBD
                     /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV
                     PALWQLQ"
     terminator      2308..2355
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    2403..2991
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2874..2885)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3008..3129
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3136..3591)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3618..3722
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3724..4081
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3932..4067
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4116..4910
                     /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    5142..5189
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
       61 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
      121 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
      181 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
      241 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
      301 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
      361 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
      421 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
      481 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
      541 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
      601 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
      661 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
      721 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
      781 ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt
      841 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      901 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      961 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     1021 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     1081 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     1141 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     1201 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     1261 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     1321 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     1381 gtttagtgaa ccgtcagatc cgctagctcg cgagtcgacg gcggccgcga attcctcgag
     1441 ggctcttcct gcatcacggg agatgcacta gttgccctac ccgagggcga gtcggtacgc
     1501 atcgccgaca tcgtgccggg tgcgcggccc aacagtgaca acgccatcga cctgaaagtc
     1561 cttgaccggc atggcaatcc cgtgctcgcc gaccggctgt tccactccgg cgagcatccg
     1621 gtgtacacgg tgcgtacggt cgaaggtctg cgtgtgacgg gcaccgcgaa ccacccgttg
     1681 ttgtgtttgg tcgacgtcgc cggggtgccg accctgctgt ggaagctgat cgacgaaatc
     1741 aagccgggcg attacgcggt gattcaacgc agcgcattca gcgtcgactg tgcaggtttt
     1801 gcccgcggga aacccgaatt tgcgcccaca acctacacag tcggcgtccc tggactggtg
     1861 cgtttcttgg aagcacacca ccgagacccg gacgcccaag ctatcgccga cgagctgacc
     1921 gacgggcggt tctactacgc gaaagtcgcc agtgtcaccg acgccggcgt gcagccggtg
     1981 tatagccttc gtgtcgacac ggcagaccac gcgtttatca cgaacgggtt cgtcagccac
     2041 gctactggcc tcaccggtct gaactcaggc ctcacgacaa atcctggtgt atccgcttgg
     2101 caggtcaaca cagcttatac tgcgggacaa ttggtcacat ataacggcaa gacgtataaa
     2161 tgtttgcagc cccacacctc cttggcagga tgggaaccat ccaacgttcc tgccttgtgg
     2221 cagcttcaat gactgcagga aggggatccg gctgctaaca aagcccgaaa ggaagctgag
     2281 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     2341 ttgaggggtt ttttgctgaa aggaggaact atatccggag gagacaatac cggaatctag
     2401 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     2461 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     2521 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     2581 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     2641 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     2701 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     2761 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     2821 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     2881 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     2941 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcaattgt
     3001 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3061 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3121 tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3181 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3241 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3301 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3361 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3421 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3481 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3541 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3601 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3661 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3721 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3781 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3841 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3901 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3961 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4021 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4081 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4141 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4201 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4261 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     4321 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4381 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4441 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4501 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4561 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4621 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     4681 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4741 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4801 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4861 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4921 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4981 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5041 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     5101 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     5161 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     5221 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     5281 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     5341 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     5401 ttaatttaa
//
