LOCUS       sequence-324723-        6716 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To generate site specific attP-GA integrant cell lines upon 
            co-delivery with BxbI. Contains IRES(Myb)-T7-Halo-PDGFR_TM-T2A-Puro.
            .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6716)
  TITLE     Huang Lab Starport System Plasmids
REFERENCE   2  (bases 1 to 6716)
  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: StarShip02
FEATURES             Location/Qualifiers
     source          1..6716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1028..1065
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     RBS             1333..1344
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1351..2064
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2076..2147
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2163..2190
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             2473..2505
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             2512..3399
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="AEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     CDS             3571..3624
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3631..4227
                     /codon_start=1
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    4236..4370
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4260..4279)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4314..4333
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4495..4511)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4495..4511)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4508..4530)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4519..4535
                     /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(4543..4549,4550..4567,4568..4573))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4588..4609
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4726..4743)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4897..5485)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4977..4996)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5656..6447,6448..6516))
                     /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     6279..6298
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6517..6621)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6689..6707
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagactgtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gacgcgtatt gggatcactg
      421 cagatcgaat tcgagttggc gcgcctgtca ttctaaatct ctctttcagc ctaaagcttt
      481 ttccccgtat ccccccaggt gtctgcaggc tcaaagagca gcgagaagcg ttcagaggaa
      541 agcgatcccg tgccaccttc cccgtgcccg ggctgtcccc gcacgctgcc ggctcgggga
      601 tgcgggggga gcgccggacc ggagcggagc cccgggcggc tcgctgctgc cccctagcgg
      661 gggagggacg taattacatc cctgggggct ttgggagggg gctgtccccg tgagctccca
      721 ggcgcgcctg tcattctaaa tctctctttc agcctaaagc tttttccccg tatcccccca
      781 ggtgtctgca ggctcaaaga gcagcgagaa gcgttcagag gaaagcgatc ccgtgccacc
      841 ttccccgtgc ccgggctgtc cccgcacgct gccggctcgg ggatgcgggg ggagcgccgg
      901 accggagcgg agccccgggc ggctcgctgc tgccccctag cgggggaggg acgtaattac
      961 atccctgggg gctttggggg ggggctgtcc ccgtgagctc aataaaagag cccactacaa
     1021 acgggccggc ttgtcgacga cggcggactc cgtcgtcagg atcatccggg agctttgcgg
     1081 gttgccattt gtaactccgg atcctaaaat tcctgtcctg ttctctgtct cttctaggtt
     1141 gggggccgtc ccgctcctaa ggcaggaata tgtgagacca gaccgaaagt gaaacgtgat
     1201 ttcatgcgtc attttgaaca ttttgtaaat cttatttaat aatgtgtgcg gcaattcaca
     1261 tttaatttat gaatgttttc ttaacatcgc ggcaactcaa gaaacggcag gttcggatct
     1321 tagctactag agaaagagga gaaatactag atgcgtaaag gcgaagagct gttcactggt
     1381 gtcgtcccta ttctggtgga actggatggt gatgtcaacg gtcataagtt ttccgtgcgt
     1441 ggcgagggtg aaggtgacgc aactaatggt aaactgacgc tgaagttcat ctgtactact
     1501 ggtaaactgc cggttccttg gccgactctg gtaacgacgc tgacttatgg tgttcagtgc
     1561 tttgctcgtt atccggacca tatgaagcag catgacttct tcaagtccgc catgccggaa
     1621 ggctatgtgc aggaacgcac gatttccttt aaggatgacg gcacgtacaa aacgcgtgcg
     1681 gaagtgaaat ttgaaggcga taccctggta aaccgcattg agctgaaagg cattgacttt
     1741 aaagaggacg gcaatatcct gggccataag ctggaataca attttaacag ccacaatgtt
     1801 tacatcaccg ccgataaaca aaaaaatggc attaaagcga attttaaaat tcgccacaac
     1861 gtggaggatg gcagcgtgca gctggctgat cactaccagc aaaacactcc aatcggtgat
     1921 ggtcctgttc tgctgccaga caatcactat ctgagcacgc aaagcgttct gtctaaagat
     1981 ccgaacgaga aacgcgatca tatggttctg ctggagttcg taaccgcagc gggcatcacg
     2041 catggtatgg atgaactgta caaatgacca ggcatcaaat aaaacgaaag gctcagtcga
     2101 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca
     2161 ctggctcacc ttcgggtggg cctttctgcg tttataggtc tcatggctag atatcaacct
     2221 gtttcctcct cctccttctc ctcctcctcc gtgacctcct cctcctcttt ctcctgagaa
     2281 acttcgcccc agcggtgcgg agcgccgctg cgcagccggg gagggacgca ggcaggcggc
     2341 gggcagcggg aggcggcagc ccggtgcggt ccccgcggct ctcggcggag ccccgcgccc
     2401 gccgcgccta tggagactga caccctcttg ctctgggtcc tgctgctgtg ggttccgggt
     2461 agtaccggtg acatggctag catgactggt ggacagcaaa tgggtggttc tgcagaaatc
     2521 ggtactggct ttccattcga cccccattat gtggaagtcc tgggcgagcg catgcactac
     2581 gtcgatgttg gtccgcgcga tggcacccct gtgctgttcc tgcacggtaa cccgacctcc
     2641 tcctacgtgt ggcgcaacat catcccgcat gttgcaccga cccatcgctg cattgctcca
     2701 gacctgatcg gtatgggcaa atccgacaaa ccagacctgg gttatttctt cgacgaccac
     2761 gtccgcttca tggatgcctt catcgaagcc ctgggtctgg aagaggtcgt cctggtcatt
     2821 cacgactggg gctccgctct gggtttccac tgggccaagc gcaatccaga gcgcgtcaaa
     2881 ggtattgcat ttatggagtt catccgccct atcccgacct gggacgaatg gccagaattt
     2941 gcccgcgaga ctttccaggc cttccgcacc accgacgtcg gccgcaagct gatcatcgat
     3001 cagaacgttt ttatcgaggg tacgctgccg atgggtgtcg tccgcccgct gactgaagtc
     3061 gagatggacc attaccgcga gccgttcctg aatcctgttg accgcgagcc actgtggcgc
     3121 ttcccaaacg agctgccaat cgccggtgag ccagcgaaca tcgtcgcgct ggtcgaagaa
     3181 tacatggact ggctgcacca gtcccctgtc ccgaagctgc tgttctgggg caccccaggc
     3241 gttctgatcc caccggccga agccgctcgc ctggccaaaa gcctgcctaa ctgcaaggct
     3301 gtggacatcg gcccgggtct gaatctgctg caagaggaca acccggacct gatcggcagc
     3361 gagatcgcgc gctggctgtc gacgctcgag atttccggcg gttctaacgc cgtgggccag
     3421 gatacgcagg aggtgatcgt cgtgccccac agcttgccct tcaaggtcgt ggttattagt
     3481 gctatcctgg cgttggtggt gctgactatt atctccctta tcatacttat catgctgtgg
     3541 caaaaaaaac cccgaggatc cggaagcgga gagggcagag gaagtctgct aacatgcggt
     3601 gacgtcgagg agaatcctgg acctggtggc accgagtaca agcctaccgt gcgcctggcc
     3661 actcgcgatg atgtgccccg cgccgtccgc actctggccg ccgctttcgc cgactacccc
     3721 gctacccggc acaccgtgga ccccgaccgg cacatcgagc gtgtgacaga gttgcaggag
     3781 ctgttcctga cccgcgtcgg gctggacatc ggcaaggtgt gggtagccga cgacggcgcg
     3841 gccgtggccg tgtggactac ccccgagagc gttgaggccg gcgccgtgtt cgccgagatc
     3901 ggcccccgaa tggccgagct gagcggcagc cgcctggccg cccagcagca aatggagggc
     3961 ctgcttgccc cccatcgtcc caaggagcct gcctggtttc tggccactgt aggagtgagc
     4021 cccgaccacc agggcaaggg cttgggcagc gccgtcgtgt tgcccggcgt agaggccgcc
     4081 gaacgcgccg gtgtgcccgc ctttctcgaa acaagcgcac caagaaacct tccattctac
     4141 gagcgcctgg gcttcaccgt gaccgccgat gtcgaggtgc ccgagggacc taggacctgg
     4201 tgtatgacac gaaaacctgg cgcctaagcg gccgcgatcc agacatgata agatacattg
     4261 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     4321 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     4381 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     4441 aaaacctcta caaatggctg atcccaatgg cgcgccgagc ttggcgtaat catggtcata
     4501 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4561 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4621 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4681 acgcgcgggg agaggcggtt tgcgtattgg gcgctgttcc gcttcctcgc tcactgactc
     4741 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4801 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4861 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4921 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4981 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5041 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5101 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5161 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5221 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5281 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5341 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5401 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5461 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5521 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5581 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5641 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5701 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5761 cttaccatct ggccccagtg ctgcaatgat accgcgagaa ccacgctcac cggctccaga
     5821 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5881 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5941 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6001 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6061 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6121 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6181 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6241 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6301 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6361 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6421 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6481 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6541 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6601 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     6661 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct tttgtc
//
