LOCUS       sequence-137346-        5913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression for structure determination; may not be full 
            ORF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5913)
  TITLE     SGC Oxford
REFERENCE   2  (bases 1 to 5913)
  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"
COMMENT     Sequence Label: SPHK1
FEATURES             Location/Qualifiers
     source          1..5913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..92
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     17..34
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             154..171
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             196..216
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     polyA_signal    1500..1634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1537..1556)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1591..1610
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1663..1828
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2012..2467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2099..2118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2309..2330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2494..2598
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2599..2667,2668..3459)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2817..2836)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3630..4218
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4119..4138
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4372..4389
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4521..4745
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4812..5345)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5534..5539,5540..5556,5557..5562))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 atcatggaga taattaaaat gataaccatc tcgcaaataa ataagtattt tactgttttc
       61 gtaacagttt tgtaataaaa aaacctataa atattccgga ttattcatac cgtcccacca
      121 tcgggcgcgg atctcggtcc gaaaaccatg ggccaccatc atcatcatca ttcttctggt
      181 gtagatctgg gtaccgagaa cctgtacttc caatccatgg agccgcgggt cgaggttatg
      241 gatccagcgg gcggcccccg gggcgtgctc ccgcggccct gccgcgtgct ggtgctgctg
      301 aacccgcgcg gtggcaaggg caaggccttg cagctcttcc ggagtcacgt gcagcccctt
      361 ttggctgagg ctgaaatctc cttcacgctg atgctcactg agcggcggaa ccacgcgcgg
      421 gagctggtgc ggtcggagga gctgggccgc tgggacgctc tggtggtcat gtctggagac
      481 gggctgatgc acgaggtggt gaacgggctc atggagcggc ctgactggga gaccgccatc
      541 cagaagcccc tgtgtagcct cccagcaggc tctggcaacg cgctggcagc ttccttgaac
      601 cattatgctg gctatgagca ggtcaccaat gaagacctcc tgaccaactg cacgctattg
      661 ctgtgccgcc ggctgctgtc acccatgaac ctgctgtctc tgcacacggc ttcggggctg
      721 cgcctcttct ctgtgctcag cctggcctgg ggcttcattg ctgatgtgga cctagagagt
      781 gagaagtatc ggcgtctggg ggagatgcgc ttcactctgg gcaccttcct gcgtctggca
      841 gccctgcgca cctaccgcgg ccgactggcc tacctccctg taggaagagt gggttccaag
      901 acacctgcct cccccgttgt ggtccagcag ggcccggtag atgcacacct tgtgccactg
      961 gaggagccag tgccctctca ctggacagtg gtgcccgacg aggactttgt gctagtcctg
     1021 gcactgctgc actcgcacct gggcagtgag atgtttgctg cacccatggg ccgctgtgca
     1081 gctggcgtca tgcatctgtt ctacgtgcgg gcgggagtgt ctcgtgccat gctgctgcgc
     1141 ctcttcctgg ccatggagaa gggcaggcat atggagtatg aatgccccta cttggtatac
     1201 gtgcccgtgg tcgccttccg tttggagccc aaggatggga aaggtgtgtt tgcagtggat
     1261 ggggaattga tggttagcga ggccgtgcag ggccaggtgc acccaaacta cttctggatg
     1321 gtcagctgac agtaaaggtg gatacggatc cgaattcgag ctccgtcgac aagcttgtcg
     1381 agaagtacta gaggatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     1441 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     1501 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     1561 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     1621 atcatgtctg gatctgatca ctgatatcgc ctaggagatc cgaaccagat aagtgaaatc
     1681 tagttccaaa ctattttgtc atttttaatt ttcgtattag cttacgacgc tacacccagt
     1741 tcccatctat tttgtcactc ttccctaaat aatccttaaa aactccattt ccacccctcc
     1801 cagttcccaa ctattttgtc cgcccacagc ggggcatttt tcttcctgtt atgtttttaa
     1861 tcaaacatcc tgccaactcc atgtgacaaa ccgtcatctt cggctacttt ttctctgtca
     1921 cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga ctgaatatca
     1981 acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     2041 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     2101 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2161 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2221 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     2281 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2341 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     2401 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     2461 acaatttcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2521 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2581 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     2641 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     2701 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     2761 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     2821 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     2881 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     2941 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3001 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3061 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3121 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3181 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3241 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3301 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3361 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3421 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     3481 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3541 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3601 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3661 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3721 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     3781 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3841 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3901 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3961 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4021 cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4081 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4141 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4201 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4261 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4321 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     4381 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     4441 atttcacacc gcagaccagc cgcgtaacct ggcaaaatcg gttacggttg agtaataaat
     4501 ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa caaaatagat
     4561 ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga aatcagtcca
     4621 gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact cttcattttc
     4681 tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg gtaaagacta
     4741 tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc gatctcggct
     4801 tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac ttcttcccgt
     4861 atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc ttgcacgtag
     4921 atcacataag caccaagcgc gttggcctca tgcttgagca gattgatgag cgcggtggca
     4981 atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat ctcactacgc
     5041 ggctgctcaa acctgggcag aacgtaagcc gcgagagcgc caacaaccgc ttcttggtcg
     5101 aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta atcggagtcc
     5161 ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag atcaagagca
     5221 gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat gcccatactt
     5281 gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt gctgctgcgt
     5341 aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt
     5401 ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga aaaccgccac
     5461 tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatacg
     5521 ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc gtgccttcat
     5581 ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg aggcatttct
     5641 gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg cattggcggc
     5701 cttgctgttc ttctacggca aggtgctgtg cacggatctg ccctggcttc aggagatcgg
     5761 aagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag tggttcgcat
     5821 cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct atagttctag
     5881 tggttggcta cgtatactcc ggaatattaa tag
//
