LOCUS       sequence-232223-        6518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hansenula polymorpha expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6518)
  TITLE     pHIPH4
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6518)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pHIPH4
FEATURES             Location/Qualifiers
     source          1..6518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1629..1648)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1630..1648)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1658..1675)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1658..1674)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1667..1689)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1816..2271
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1903..1922)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2113..2134
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2297..2401
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2402..2470,2471..3262)
                     /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(2620..2639)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3433..4021
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3922..3941
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4175..4192
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4309..4330
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4345..4350,4351..4368,4369..4375)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4383..4399
                     /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)."
     primer_bind     4388..4410
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4407..4423
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4407..4423
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4442..4462
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4444..4462
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      complement(4542..4739)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(4745..5773)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             complement(4745..5773)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(5792..5839)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(5846..6259)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 atggcgtagg aaggctggat gtccaggccc ggggatcgat cctctagagt cgacctgcag
       61 gcatgcaagc ttgtttttgt actttagatt gatgtcacca ccgtgcactg gcagcagtat
      121 ttatagatgg accgtgtggg gacggttggg tacacttagc ggcagcgctg accccatctg
      181 tgatcaagta gggcaaaaac tggggatgtc ggagtcactg cacggtagca taagaattta
      241 ctttctggcc ggttcacccg catttgcact gtggagaaac agcctgtccg acaccccacc
      301 agttgccaca tcggccctct gctgctctgg tgattttctg gtagcaggca cagacagcag
      361 tgggtagcgc cgtccggtta ggcaaggtca cgttgtaggc tacgccagca aacagagcct
      421 cacatgacac catccagctg cgtcctcgaa gcgaaaagtt cggttgcggc tgcagaaccc
      481 cctcagttgc cagattcaca agttttacgc gacggctaaa gcgagtgggt tttaaaaact
      541 tgcggtgcaa ggatgcatgc ggcaacaatt aattggtgca tccagcacag caagcccagt
      601 ctcgagatgt ccagtcgcta cagagtggag tacgcactca aggaacaccg tcgagatggc
      661 ctcatagaat ggatcaaggg cctgctggcc acgccgttcg tcctgtacgc ggtgaagagc
      721 aacggcatct ctgcagtgga cgacctcatg gtaaactctg aggcaaaacg ccgctacgcg
      781 gaaatcttcc acgacctcga actcctcatc gacgacaaca ttgaaatgac caaagccggc
      841 acccccgaat tgtctcggct cgtgcagctg gttccgagcg ttggcagctt cttcacgaga
      901 ctgcctctgg aaaaggcctt ctacatcgag gacgagcgcc gcgccatcag caaacgccgg
      961 cttgtggccc cctcgttcaa cgacgtccgg ctcattctca acacggccca gctgttggag
     1021 atgtcgcggt tcttccattc caaaaccatc cgagatcgca agctgcagct cattacattc
     1081 gatggtgaca tcacactgta cgacgacggc aaaaatttcg atgccgagtc gcccatcctg
     1141 ccccacctca tcaaactaat ggccaaggac ctctatgtgg gtatcgtcac cgcggccggc
     1201 tacagcgacg cgaacaagta ctacgagcgc ctcaagggcc tcatcgacgc cgtccagacg
     1261 tccccgctgc tcacaggcca ccagaaagag aacctgttca ttatgggcgg cgaggcaaac
     1321 tacctcttcc ggtacagtaa cgaggagcag agattacgct tctactccaa agacagatgg
     1381 ctgctcgaga acatgctgaa ttggtccgag gaggacattc atctgacact ggactttgcg
     1441 caggacgttc taaacgacct cgttcacaaa ctgggctcgc cagccaccgt ggtccgcaag
     1501 gagcgtgccg tcggcctggt tccattaccg ggccacaagc tgatccgcga gcagctcgag
     1561 gagatcgttc tccgcgtcga gatccgatat cgccgtggcg gccgccaccg cggtggagct
     1621 ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc
     1681 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     1741 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     1801 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     1861 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     1921 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     1981 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     2041 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2101 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     2161 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     2221 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     2281 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2341 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2401 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     2461 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     2521 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     2581 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     2641 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     2701 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     2761 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2821 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     2881 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     2941 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3001 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3061 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3121 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     3181 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3241 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3301 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3361 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3421 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3481 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3541 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3601 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3661 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3721 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3781 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3841 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3901 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3961 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4021 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4081 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     4141 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     4201 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     4261 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     4321 gctcactcat taggcacccc aggctttaca ctttatgctc ccggctcgta tgttgtgtgg
     4381 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg
     4441 cgcaattaac cctcactaaa gggaacaaaa gctgggtaca tcatcgatga attcgagctc
     4501 gttttcgaca ctggatggcg gcgttagtat cgaatcgaca gcagtatagc gaccagcatt
     4561 cacatacgat tgacgcatga tattactttc tgcgcactta acttcgcatc tgggcagatg
     4621 atgtcgaggc gaaaaaaaat ataaatcacg ctaacatttg attaaaatag aacaactaca
     4681 atataaaaaa actatacaaa tgacaagttc ttgaaaacaa gaatcttttt attgtcagta
     4741 ctgattattc ctttgccctc ggacgagtgc tggggcgtcg gtttccacta tcggcgagta
     4801 cttctacaca gccatcggtc cagacggccg cgcttctgcg ggcgatttgt gtacgcccga
     4861 cagtcccggc tccggatcgg acgattgcgt cgcatcgacc ctgcgcccaa gctgcatcat
     4921 cgaaattgcc gtcaaccaag ctctgataga gttggtcaag accaatgcgg agcatatacg
     4981 cccggagccg cggcgatcct gcaagctccg gatgcctccg ctcgaagtag cgcgtctgct
     5041 gctccataca agccaaccac ggcctccaga agaagatgtt ggcgacctcg tattgggaat
     5101 ccccgaacat cgcctcgctc cagtcaatga ccgctgttat gcggccattg tccgtcagga
     5161 cattgttgga gccgaaatcc gcgtgcacga ggtgccggac ttcggggcag tcctcggccc
     5221 aaagcatcag ctcatcgaga gcctgcgcga cggacgcact gacggtgtcg tccatcacag
     5281 tttgccagtg atacacatgg ggatcagcaa tcgcgcatat gaaatcacgc catgtagtgt
     5341 attgaccgat tccttgcggt ccgaatgggc cgaacccgct cgtctggcta agatcggccg
     5401 cagcgatcgc atccatggcc tccgcgaccg gctgcagaac agcgggcagt tcggtttcag
     5461 gcaggtcttg caacgtgaca ccctgtgcac ggcgggagat gcaataggtc aggctctcgc
     5521 tgaattcccc aatgtcaagc acttccggaa tcgggagcgc ggccgatgca aagtgccgat
     5581 aaacataacg atctttgtag aaaccatcgg cgcagctatt tacccgcagg acatatccac
     5641 gccctcctac atcgaagctg aaagcacgag attcttcgcc ctccgagagc tgcatcaggt
     5701 cggagacgct gtcgaacttt tcgatcagaa acttctcgac agacgtcgcg gtgagttcag
     5761 gctttttacc catggtttag ttcctcacct tgtcgtatta tactatgccg atatactatg
     5821 ccgatgatta attgtcaaca ccgcccttag attagattgc tatgctttct ttctaatgag
     5881 caagaagtaa aaaaagttgt aatagaacaa gaaaaatgaa actgaaactt gagaaattga
     5941 agaccgttta ttaacttaaa tatcaatggg aggtcatcga aagagaaaaa aatcaaaaaa
     6001 aaaaattttc aagaaaaaga aacgtgataa aaatttttat tgcctttttc gacgaagaaa
     6061 aagaaacgag gcggtctctt ttttcttttc caaaccttta gtacgggtaa ttaacgacac
     6121 cctagaggaa gaaagagggg aaatttagta tgctgtgctt gggtgttttg aagtggtacg
     6181 gcgatgcgcg gagtccgaga aaatctggaa gagtaaaaaa ggagtagaaa cattttgaag
     6241 ctatggtgtg tgggggatct gaacctcgac tttctggatc agcccgccca ggccggccct
     6301 gagcccgctg ctcctgcgca ggaggatgac gccgggctga tgcctacaga ccagatggaa
     6361 aacctatttt cgcagttcga cgagctcgtc aatagtggag attactagat tgtcgtgtcc
     6421 tatatactta tttaccgcaa caagagcctt caaaggcaag tctcgaggtt ttgtccttgg
     6481 tctccttgtg cacggccctc ttagcctcgg aggtggtc
//
