LOCUS       sequence-423561-        3593 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Syn8 terminator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3593)
  AUTHORS   Jiang W, Wang S, Avila P, Jorgensen TS, Yang Z, Borodina I
  TITLE     Combinatorial iterative method for metabolic engineering of Yarrowia
            lipolytica: application for betanin biosynthesis.
  JOURNAL   Metab Eng. 2024 Sep 9:S1096-7176(24)00110-1. doi: 
            10.1016/j.ymben.2024.09.003.
  PUBMED    39260817
REFERENCE   2  (bases 1 to 3593)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Metab Eng. 
            2024 Sep 9:S1096-7176(24)00110-1. doi: 10.1016/j.ymben.2024.09.003."
COMMENT     Sequence Label: pCfB12710
FEATURES             Location/Qualifiers
     source          1..3593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(606..980)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     regulatory      977..986
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1186..1980)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(1186..1980)
                     /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"
     primer_bind     complement(1297..1316)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1907..1926
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(2329..2631)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(2431..2450)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     2754..2776
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2768..2785
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2769..2785
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2792..2811
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2792..2810
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3035..3053)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3036..3053)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3071..3087)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3071..3087)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3084..3106)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3095..3111
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3119..3125,3126..3143,3144..3149))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3164..3185
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3302..3319)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(3473..3593,1..468))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3553..3572)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
       61 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
      121 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
      181 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
      241 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
      301 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
      361 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
      421 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
      481 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
      541 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttagc
      601 acgtgtcagt cctgctcctc ggccacgaag tgcacgcagt tgccggccgg gtcgcgcagg
      661 gcgaactccc gcccccacgg ctgctcgccg atctcggtca tggccggccc ggaggcgtcc
      721 cggaagttcg tggacacgac ctccgaccac tcggcgtaca gctcgtccag gccgcgcacc
      781 cacacccagg ccagggtgtt gtccggcacc acctggtcct ggaccgcgct gatgaacagg
      841 gtcacgtcgt cccggaccac accggcgaag tcgtcctcca cgaagtcccg ggagaacccg
      901 agccggtcgg tccagaactc gaccgctccg gcgacgtcgc gcgcggtgag caccggaacg
      961 gcactggtca acttggccat ggtggccctc ctcacgtgct attattgaag catttatcag
     1021 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     1081 gttccgcgca catttccccg aaaagtgcca cctgatgcgg tgtgaaatac cgcacagatg
     1141 cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta ataattcaga agaactcgtc
     1201 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag
     1261 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat
     1321 gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc
     1381 attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc
     1441 gtcgggcatg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc
     1501 ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat
     1561 gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg
     1621 cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc
     1681 ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag
     1741 cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg
     1801 cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc
     1861 tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc
     1921 gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat
     1981 gcgaaacgat cctcatcctg tctcttgatc agagcttgat cccctgcgcc atcagatcct
     2041 tggcggcaag aaagccatcc agtttacttt gcagggcttc ccaaccttac cagagggcgc
     2101 cccagctggc aattccggtt cgcttgctgt ccataaaacc gcccagtcta gctatcgcca
     2161 tgtaagccca ctgcaagcta cctgctttct ctttgcgctt gcgttttccc ttgtccagat
     2221 agcccagtag ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctacgt
     2281 gaaaaggatc taggtgaaga tcctttttga taatctcatg cctgacattt atattcccca
     2341 gaacatcagg ttaatggcgt ttttgatgtc attttcgcgg tggctgagat cagccacttc
     2401 ttccccgata acggagaccg gcacactggc catatcggtg gtcatcatgc gccagctttc
     2461 atccccgata tgcaccaccg ggtaaagttc acgggagact ttatctgaca gcagacgtgc
     2521 actggccagg gggatcacca tccgtcgccc cggcgtgtca ataatatcac tctgtacatc
     2581 cacaaacaga cgataacggc tctctctttt ataggtgtaa accttaaact gccgtacgta
     2641 taggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2701 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     2761 acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg gcgaattggg
     2821 ccctctagat gcatgctcga gcggccgcca gtgtgatgga tatctgcaga attcgccctt
     2881 ggtctctact caagctttga aagatgatac tctttattcc tacataagta aatgagttta
     2941 tatatagaag agaccaaggg cgaattccag cacactggcg gccgttacta gtggatccga
     3001 gctcggtacc aagcttgatg catagcttga gtattctata gtgtcaccta aatagcttgg
     3061 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3121 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3181 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3241 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3301 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3361 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3421 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3481 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3541 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgc
//
